Protein kinase B inhibitors

ABSTRACT

The invention relates to a novel group of compounds of Formula (I) or salts thereof:wherein Y, Z1, Z2, R1, R4, R5 and n are as described in the specification, which may be useful in the treatment or prevention of a disease or medical condition mediated through protein kinase B (PKB) such as cancer. The invention also relates to pharmaceutical compositions comprising said compounds, methods of treatment of diseases mediated by PKB using said compounds and methods for preparing compounds of Formula (I).

This application is a Continuation of U.S. application Ser. No.16/841,820 filed on 7 Apr. 2020, which is a Continuation of U.S.application Ser. No. 16/028,979 filed 6 Jul. 2018, (now granted as U.S.Pat. No. 10,654,855 on 19 May 2020), which is a Continuation of U.S.application Ser. No. 15/351,481 filed on 15 Nov. 2016 (now granted asU.S. Pat. No. 10,059,714 on 28 Aug. 2018), which is a Continuation ofU.S. application Ser. No. 14/626,303, filed on 19 Feb. 2015 (now grantedas U.S. Pat. No. 9,492,453 on Nov. 15, 2016), which is a Continuation ofU.S. application Ser. No. 13/324,191, filed on 13 Dec. 2011 (Abandoned),which is a Continuation of U.S. application Ser. No. 12/249,477, filedon 10 Oct. 2008 (now granted as U.S. Pat. No. 8,101,623 on 24 Jan.2012), which claims the benefit under 35 U.S.C. § 119(e) of ApplicationNo. 60/979,192 (US) filed on 11 Oct. 2007 and Application No. 61/047,862(US) filed on 25 Apr. 2008, each of which is hereby incorporated hereinby reference in their entireties.

The present invention relates to a novel group of bicyclic heterocycleswhich may be useful in the treatment or prevention of a disease ormedical condition mediated through protein kinase B (PKB, also known asAKT). Such compounds may therefore be useful in the treatment orprevention of a number of different cancers. The invention also relatesto pharmaceutical compositions comprising said compounds, to processesfor the manufacture of said compounds and to methods of treatment ofdiseases mediated by PKB using said compounds.

PKB is a component of the phosphatidyl 3-kinase (PI3K) signallingpathway which plays an important part in cell proliferation andsurvival, including cellular responses to growth factors. Upon bindingof a growth factor, for example epidermal growth factor (EGF), to itscell surface receptor tyrosine kinase, for example EGF receptor (EGFR),the receptor dimerises and undergoes autophosphorylation. Thisautophosphorylation event allows the 85 kDa regulatory subunit of PI3K(p85) to interact with the receptor either directly or via an adaptorprotein, for example growth factor receptor-bound protein 2 (GRB2), andthereby activate the 110 kDa catalytic subunit of PI3K (p110). Uponactivation, p110 catalyses the phosphorylation ofphosphatidylinositol-4,5-bisphosphate (PIP₂) to producephosphatidylinositol-3,4,5-triphosphate (PIP₃), a second messengermolecule that recruits both phosphatidylinositol-dependent kinase 1(PDK1) and PKB to the plasma membrane where PDK1 phosphorylates andactivates PKB.

There are three known isoforms of PKB (PKBα/AKT1, PKBβ/AKT2 andPKBγ/AKT3), derived from three distinct genes. Activation of PKBα isassociated with cell signalling events that mediate cell proliferationand survival, whereas activation of PKBβ is associated with invasion,motility and insulin-mediated metabolic processes. Activated PKBprotects cells from apoptosis by inactivating proapoptotic factors, forexample the BAD, procaspase-9 and forkhead (FKHR) transcription factors,and activating transcription factors that upregulate antiapoptoticgenes, for example cyclic-AMP response element binding protein (CREB).PKB can also contribute to cell survival by inactivation of p53 viaphosphorylation of MDM2. Similarly, activated PKB induces cellproliferation by activating proteins involved in cell growth andmetabolism, for example by a pathway leading to activation of themammalian target of rapamycin (mTOR) and via glycogen synthase kinase-3(GSK3).

PKB-mediated stimulation of cell proliferation and protection fromapoptosis therefore favour tumourigenesis and genetic disturbances ofcomponents within the PI3K pathway are commonly found in cancer. Forexample, mutation or amplification of the genes encoding the p110isoforms of PI3K are found in breast cancers, bowel cancer, ovariancancer, head and neck and cervical squamous cancers, gastric and lungcancers, angioplastic oligodendrogliomas, amaplastic astrocytomas,glioblastoma multiforme and medulloblastomas. Similarly, mutationamplification and/or overexpression of the genes encoding the PKBisoforms are found in pancreatic, breast and ovarian tumours.Furthermore, the gene encoding for PTEN (a phosphatase which has areverse role to PI3K, catalysing the conversion of PIPS to PIP₂) isinactivated in many tumour types, including ovarian, colorectal, breast,glioma, melanoma, lung, leukaemias and lymphomas; this results inactivation of PKB/AKT.

In view of the importance of the PI3K signalling pathway in tumour cellproliferation and survival, any compound that disrupted this pathway,including PKB inhibitors, may be useful in the treatment of cancer.Detailed reviews of the PI3K signalling pathway and its involvement intumourigenisis are provided by Hennessy et al., Nature Reviews/DrugDiscovery (December 2005) Vol. 4, 988-1004, and Cully et al., Naturereviews/Cancer (March 2006) Vol. 6, 184-192.

The voltage-dependent potassium channel encoded by the humanether-a-go-go-related gene (hERG) is believed to play a key role inrepolarisation of the ventricular cardiac action potential. Changes inits activity, caused either by inherited mutations of the gene sequenceor pharmacological modification, can lead to prolongation of actionpotential duration. This can lead to prolongation of the QT intervalrecorded in man on an electrocardiogram and to a potentially fatalcardiac arrhythmia known as Torsades de Pointes (Vandenberg et al.(2001). Trends Pharmacol. Sci. 22, 240-246). Recent regulatoryguidelines (CPMP/ICH/539/00) recommend that an in vitro assayinvestigating the effects of test compounds at the hERG channel could beone element of a pre-clinical strategy aiming to predict the likelihoodthat new chemical entities will prolong the QT interval recorded in manon an electrocardiogram. As such, the elimination of hERG blockingactivity remains an important consideration in the development of anynew drug.

A number of compounds have been described that target the PI3K pathway.For example WO2006/046023 and WO2006/046024 (Astex Therapeutics Limited)describe purine, purinone and deazapurinone compounds that inhibit ormodulate the activity of protein kinase B (PKB) and protein kinase A(PKA). However, there still exists the need for further improved agentshaving superior potency against PKB and/or advantageous physicalproperties (for example, higher aqueous solubility, higher permeability,and/or lower plasma protein binding) and/or favourable toxicity profiles(for example a descreased hERG blocking liability) and/or favourablemetabolic profiles in comparison with other known PKB inhibitors.

The applicants have surprisingly found that certain bicyclic heterocyclederivatives are particularly effective at inhibiting PKB activity andmay therefore be useful in the treatment of disease states in which PKBactivity is implicated, for example cancer.

According to a first aspect of the invention, there is thereforeprovided a compound of Formula (I), or a pharmaceutically acceptablesalt thereof:

wherein:

-   -   Y represents CH or N;    -   Z¹—Z² represents a group selected from C(R⁶)═CH, N═CH and        C(R⁶)═N; where        -   R⁶ represents hydrogen, fluoro, chloro, bromo, cyano,            methyl, ethyl, difluoromethyl, trifluoromethyl or            cyclopropyl;    -   n is 0, 1 or 2;    -   R¹ represents C₁₋₄alkyl, C₂₋₄alkenyl, C₂₋₄alkynyl, C₁₋₄alkoxy,        C₁₋₄alkoxyC₁₋₄alkyl, fluoroC₁₋₄alkyl, aminoC₁₋₄alkyl,        hydroxyC₁₋₄alkyl, cyano, cyanoC₁₋₄alkyl, C₃₋₆cycloalkyl,        —(CH₂)_(p)NHCOCH₃, —(CH₂)_(p)NHSO₂CH₃, —(CH₂)_(p)NHCONH₂,        —(CH₂)_(p)NHCONR²R³, —(CH₂)_(p)NR²R³, —(CH₂)_(p)SO₂NH₂,        —(CH₂)_(p)SO₂NR²R³, —(CH₂)_(p)CONH₂, —(CH₂)_(p)CONR²R³ or        —(CH₂)_(p)—R⁷; where        -   p is 0, 1, 2 or 3;        -   R² represents hydrogen or C₁₋₃alkyl;        -   R³ represents C₁₋₃alkyl; and        -   R⁷ represents phenyl;        -   R⁷ represents a 5 or 6 membered monocyclic heteroaryl ring            which comprises 1, 2 or 3 heteroatoms selected from O, N or            S; or        -   R⁷ represents a monocyclic 4, 5, or 6 membered heterocyclic            ring which comprises 1, 2 or 3 heteroatoms selected from O,            N or S;        -   wherein R⁷ is optionally substituted by 1 or 2 substituents            selected from C₁₋₄alkyl, trifluoromethyl, C₁₋₄alkoxy,            fluoro, chloro, bromo, and cyano;    -   R⁴ represents hydrogen, fluoro, chloro, bromo, cyano or        trifluoromethyl; and    -   R⁵ represents hydrogen, fluoro, chloro or bromo.

In one embodiment of the invention, there is provided a compound ofFormula (I), or a pharmaceutically acceptable salt thereof:

wherein:

-   -   Y represents CH or N;    -   Z¹—Z² represents a group selected from CH═CH, N═CH and CH═N;    -   n is 0, 1 or 2; and    -   R¹ represents C₁₋₄alkyl, aminoC₁₋₄alkyl, hydroxyC₁₋₄alkyl,        —(CH₂)_(p)NHCOCH₃, —(CH₂)_(q)NR²R³ or C₃₋₆cycloalkyl; where        -   R² represents hydrogen or C₁₋₃alkyl;        -   R³ represents C₁₋₃alkyl; and        -   p and q independently represent 2 or 3.

In a further embodiment of the invention, there is therefore provided acompound of Formula (I), or a pharmaceutically acceptable salt thereof:

wherein:

-   -   Y represents CH or N;    -   Z¹—Z² represents a group selected from CH═CH, N═CH and CH═N;    -   R¹ represents C₁₋₄alkyl, aminoC₁₋₄alkyl or C₃₋₆cycloalkyl; and    -   n is 0, 1 or 2.

The term “C₁₋₄alkyl” is intended to mean a saturated carbon chain of 1to 4 carbon atoms in length which may be straight-chained or branched.However references to individual alkyl groups such as “propyl” arespecific for the straight chain version only and references toindividual branched-chain alkyl groups such as t-butyl are specific forthe branched chain version only. For example, “C₁₋₄alkyl” includesmethyl, ethyl, propyl, isopropyl and t-butyl.

The term “C₂₋₄alkenyl” is intended to mean an unsaturated carbon chainof 2 to 4 carbon atoms in length, which may be straight-chained orbranched, containing at least one carbon to carbon double bond. Howeverreferences to individual alkenyl groups such as “propenyl” are specificfor the straight chain version only and references to individualbranched-chain alkyl groups such as tert-butenyl are specific for thebranched chain version only. For example, “C₂₋₄alkenyl” includes, but isnot limited to, ethenyl, propenyl, isopropenyl, butenyl andtert-butenyl.

The term “C₂₋₄alkynyl” is intended to mean an unsaturated carbon chainof 2 to 4 carbon atoms in length, which may be straight-chained orbranched, containing at least one carbon to carbon triple bond. Howeverreferences to individual alkynyl groups such as “propynyl” are specificfor the straight chain version only and references to individualbranched-chain alkyl groups such as tert-butynyl are specific for thebranched chain version only. For example, “C₂₋₄alkynyl” includes, but isnot limited to, ethynyl, propynyl, isopropynyl, butynyl andtert-butynyl.

The term “C₁₋₄ alkoxy” is intended to mean a saturated carbon chain of 1to 4 carbon atoms in length, which may be straight-chained or branched,linked to oxygen. For example, “C₁₋₆ alkoxy” includes, but is notlimited to, methoxy, ethoxy, propoxy and butoxy.

The term “C₁₋₄alkoxyC₁₋₄alkyl” is intended to mean a saturated carbonchain of 1 to 4 carbon atoms in length, which may be straight-chained orbranched, linked via oxygen to another saturated carbon chain of 1 to 4carbon atoms in length, which may be straight-chained or branched. Forexample, “C₁₋₄alkoxyC₁₋₄alkyl” includes, but is not limited to,methoxyethyl, methoxypropyl, ethoxypropyl, propoxyethyl andbutoxypropyl.

The term “fluoroC₁₋₄alkyl” is intended to mean a saturated carbon chainof 1 to 4 carbon atoms in length which may be straight-chained orbranched wherein at least one of the hydrogen atoms have been replacedby fluorine. For example, “fluoroC₁₋₄alkyl” includes, but is not limitedto, fluoromethyl, fluoroethyl, fluoropropyl, fluoroisopropyl,fluorobutyl, fluoroisobutyl, fluoro-tert-butyl, trifluoromethyl,pentafluoroethyl, heptafluoropropyl and nonafluorobutyl.

The term “aminoC₁₋₄alkyl” is intended to mean a saturated carbon chainof 1 to 4 carbon atoms in length, which may be straight-chained orbranched, comprising one primary amino group. For example “aminoC₁₋₄alkyl” includes aminomethyl, aminoethyl, 2-aminopropyl, 3-aminopropyl,1-aminoisopropyl and 4-aminobutyl.

The term “hydroxyC₁₋₄alkyl” is intended to mean a saturated carbon chainof 1 to 4 carbon atoms in length, which may be straight-chained orbranched, comprising a hydroxyl group. For example “hydroxyC₁₋₄alkyl”includes hydroxymethyl, hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl,1-hydroxyisopropyl and 4-hydroxybutyl.

The term “C₃₋₆cycloalkyl” is intended to mean a saturated 3 to 6membered monocyclic carbon ring. For example “C₃₋₆cycloalkyl” includescyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

The term “heteroaryl ring” is intended to mean a 5 or 6 membered,totally unsaturated, aromatic monocyclic ring which comprises 1, 2 or 3heteroatoms independently selected from nitrogen, oxygen or sulphur,linked via a ring carbon atom or a ring nitrogen atom where a bond froma nitrogen is possible, for example no bond is possible to the nitrogenof a pyridine ring, but a bond is possible through the 1-nitrogen of apyrazole ring. Examples of 5 or 6 membered heteroaryl rings include, butare not limited to, pyrrole, furan, imidazole, triazole, tetrazole,pyrazine, pyrimidine, pyridazine, pyridine, pyrazole, isoxazole,oxazole, 1,2,4 oxadiazole, isothiazole, thiazole, 1,2,4-triazole andthiophene.

The term “heterocyclic ring” is intended to mean a 4, 5 or 6 memberedfully saturated or partially saturated monocyclic ring which comprises1, 2 or 3 heteroatoms selected from nitrogen, oxygen or sulphur linkedvia a ring carbon atom or a ring nitrogen atom. Examples of 4, 5 or 6membered heterocyclic rings include azetidine, tetrahydrofuran,tetrahydropyran, pyrroline, pyrrolidine, thiazolidine, morpholine,piperidine, piperazine, dihydropyridine, dihydropyrimidine and azepane.

In further embodiments of the invention, each of the followingdefinitions of Y, Z¹—Z², R¹, R⁴, R⁵, R⁶, n and p in paragraphs (1) to(26) hereinafter may be used individually or in combination with one ofthe other following definitions to limit the broadest definition ofFormulae (I), (IA) or (IB) as appropriate.

-   -   (1) Y represents N;    -   (2) Z¹—Z² represents CH═CH;    -   (3) Z¹—Z² represents C(C₁)═CH;    -   (4) Z¹—Z² represents C(Br)═CH;    -   (5) R¹ represents C₁₋₄alkyl;    -   (6) R¹ represents aminoC₁₋₄alkyl;    -   (7) R¹ represents hydroxyC₁₋₄alkyl;    -   (8) R¹ represents C₃₋₆cycloalkyl;    -   (9) R¹ represents C₁₋₄alkoxyC₁₋₄alkyl, fluoroC₁₋₄alkyl,        aminoC₁₋₄alkyl, hydroxyC₁₋₄alkyl, cyanoC₁₋₄alkyl,        C₃₋₆cycloalkyl, —(CH₂)_(p)NHCOCH₃, —(CH₂)_(p)NHSO₂CH₃,        —(CH₂)_(p)NHCONH₂, —(CH₂)_(p)NHCONR²R³, —(CH₂)_(p)NR²R³,        —(CH₂)_(p)SO₂NH₂,    -   (6.1) —(CH₂)_(p)CONH₂, —(CH₂)_(p)CONR²R³ or —(CH₂)_(p)—R⁷;    -   (10) R¹ represents —(CH₂)_(p)—R⁷ wherein R⁷ is selected from        phenyl, piperidinyl, piperazinyl, pyrrolidinyl, morpholinyl,        imidazolyl, isoxazolyl, pyrazolyl and thiazolyl and R⁷ is        optionally substituted by a single methyl group;    -   (11) R¹ represents hydroxyethyl;    -   (12) n is 0;    -   (13) n is 1;    -   (14) n is 1 or 2;    -   (15) n is 0 or 1;    -   (16) p is 1, 2 or 3;    -   (17) R⁴ represents chloro, bromo or cyano;    -   (18) R⁴ represents chloro, bromo;    -   (19) R⁴ represents chloro;    -   (20) R⁴ represents bromo;    -   (21) R⁵ represents hydrogen;    -   (22) R⁵ represents chloro;    -   (23) R⁶ represents hydrogen;    -   (24) R⁶ represents methyl;    -   (25) R⁶ represents difluoromethyl;    -   (26) R⁶ represents trifluoromethyl.

According to another embodiment of the invention, there is provided acompound of Formula (I), or a pharmaceutically acceptable salt thereof,which is:

-   4-amino-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (R)-4-amino-N-(1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-((4-chlorophenyl)(cyclopropyl)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(2-amino-1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-(aminomethyl)-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (R)-4-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-2-hydroxyethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   N-(3-acetamido-1-(4-chlorophenyl)propyl)-4-amino-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (R)-4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-(aminomethyl)-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(3-amino-1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (R)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (R)-4-amino-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (R)-4-(aminomethyl)-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-cyanophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N-(1-(4-chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(3-hydroxy-1-phenylpropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(9H-purin-6-yl)piperidine-4-carboxamide;-   (S)-4-(aminomethyl)-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-bromophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-4-(dimethylamino)butyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-chlorophenyl)-3-(diethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   (S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(5-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-3-(methylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(4-chlorophenyl)(phenyl)methyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[2-amino-1-(4-chlorophenyl)-2-oxoethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-1-(3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-N-[(1S)-1-(4-chlorophenyl)ethyl]piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)ethyl]-1-(5-chloro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-1-(3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-N-[(1S)-1-(4-chlorophenyl)-3-hydroxypropyl]piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)-3-hydroxypropyl]-1-(5-chloro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N-[(1S)-1-(4-chlorophenyl)ethyl]piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)-3-hydroxypropyl]-1-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)ethyl]-1-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)-3-hydroxy-3-methylbutyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-cyanophenyl)ethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(3-chlorophenyl)-3-hydroxypropyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N-{(1S)-1-[4-(trifluoromethyl)phenyl]ethyl}piperidine-4-carboxamide;-   4-amino-N-[(1R)-1-(4-bromophenyl)ethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[    1-(4-chlorophenyl)-2-phenylethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[    1-(4-fluorophenyl)ethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(4-chlorophenyl)(cyano)methyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-phenylethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[1-(4-chlorophenyl)-4-pyrrolidin-1-ylbutyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[    1-(4-chlorophenyl)-4-morpholin-4-ylbutyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[1-(4-chlorophenyl)-4-piperidin-1-ylbutyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)-4-piperidin-1-ylbutyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1R)-1-(4-chlorophenyl)-4-piperidin-1-ylbutyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)-3-(4-methylpiperazin-1-yl)propyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)-3-morpholin-4-ylpropyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)-3-piperidin-1-ylpropyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)-3-piperazin-1-ylpropyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)-3-(1H-imidazol-1-yl)propyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[(1S)-1-(4-chlorophenyl)-3-pyrrolidin-1-ylpropyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-2-sulfamoylethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-2-sulfamoylethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   N-(2-acetamido-1-(4-chlorophenyl)ethyl)-4-amino-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-2-(1H-imidazol-2-yl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-Amino-N-[1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-[1-(4-chlorophenyl)-2-(3-methylisoxazol-5-yl)ethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-2-(thiazol-2-yl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)-3-oxopropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-3-methoxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-3-sulfamoylpropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(3-amino-1-(4-chlorophenyl)-3-oxopropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-3-ureidopropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;-   4-amino-N-(1-(4-chlorophenyl)-2-cyanoethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide;    or-   4-amino-N-(1-(4-chlorophenyl)-3-(methylsulfonamido)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide.

It is to be understood that, insofar as compounds of Formula (I) definedabove exist in optically active or racemic forms by virtue of theasymmetric carbon atom, the invention includes in its definition anysuch optically active or racemic form which possesses the property ofinhibiting PKB activity. The synthesis of optically active forms may becarried out by standard techniques of organic chemistry well known inthe art, for example by synthesis from optically active startingmaterials or by resolution of a racemic form. Racemic compounds andracemic intermediates thereof are drawn herein as flat structureswhereas stereospecific compounds and stereospecific intermediatesthereof are drawn with the appropriate stereochemistry indicated.

The invention also relates to any and all tautomeric forms of thecompounds of Formula (I) which are inhibitors of PKB activity.

In one embodiment of the invention, the compound of Formula (I) has theconfiguration shown in Formula (IA):

wherein Y, Z¹, Z², R¹, R⁴, R⁵ and n are as defined hereinbefore.

In another embodiment of the invention, the compound of Formula (I) hasthe configuration shown in Formula (IB):

wherein Y, Z¹, Z², R¹, R⁴, R⁵ and n are as defined hereinbefore.

Reference herein to a compound of Formula (I) should be understood torefer equally to a compound of Formula (I), (IA) or (IB).

In one embodiment of the invention, there is provided a compound ofFormula (IA), or a pharmaceutically acceptable salt thereof:

wherein:

-   -   Y represents CH or N;    -   Z¹—Z² represents a group selected from CH═CH, N═CH and CH═N;    -   n is 0, 1 or 2; and    -   R¹ represents C₁₋₄alkyl, aminoC₁₋₄alkyl, hydroxyC₁₋₄alkyl,        —(CH₂)_(p)NHCOCH₃, —(CH₂)_(q)NR²R³ or C₃₋₆cycloalkyl; where        -   R² represents hydrogen or C₁₋₃alkyl;        -   R³ represents C₁₋₃alkyl; and        -   p and q independently represent 2 or 3.

In a further embodiment of the invention, there is therefore provided acompound of Formula (IA) or a pharmaceutically acceptable salt thereof:

wherein:

-   -   Y represents CH or N;    -   Z¹—Z² represents a group selected from CH═CH, N═CH and CH═N;    -   R¹ represents C₁₋₄alkyl, aminoC₁₋₄alkyl or C₃₋₆cycloalkyl; and        -   n is 0, 1 or 2.

In one embodiment of the invention, there is provided a compound ofFormula (I), (IA) or (IB) as defined hereinbefore, or a pharmaceuticallyacceptable salt thereof, wherein the compound of Formula (I), (IA) or(IB) is other than(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide.

A suitable pharmaceutically acceptable salt of a compound of Formula (I)is, for example, an acid-addition salt of a compound of the inventionwhich is sufficiently basic, for example, an acid-addition salt with,for example, an inorganic or organic acid, for example hydrochloric,hydrobromic, sulphuric, phosphoric, trifluoroacetic, citric or maleicacid.

It will be understood that certain compounds of the present inventionmay exist in solvated, for example hydrated, as well as unsolvatedforms. It is to be understood that the present invention encompasses allsuch solvated forms which are inhibitors of PKB activity.

The compounds of the Formula (I) may be administered in the form of aprodrug which is broken down in the human or animal body to give acompound of the Formula (I). Examples of prodrugs include in vivohydrolysable esters of a compound of the formula (I). Various forms ofprodrugs are known in the art. For examples of such prodrug derivatives,see:

-   a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and    Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et    al. (Academic Press, 1985);-   b) A Textbook of Drug Design and Development, edited by    Krogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design and    Application of Prodrugs”, by H. Bundgaard p. 113-191 (1991);-   c) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);-   d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285    (1988); and-   N. Kakeya, et al., Chem Pharm Bull, 32, 692 (1984).

According to a further aspect of the invention there is provided apharmaceutical composition, which comprises a compound of the Formula(I), or a pharmaceutically acceptable salt thereof, as definedhereinbefore in association with a pharmaceutically-acceptable diluentor carrier.

The compositions of the invention may be in a form suitable for oral use(for example as tablets, lozenges, hard or soft capsules, aqueous oroily suspensions, emulsions, dispersible powders or granules, syrups orelixirs), for topical use (for example as creams, ointments, gels, oraqueous or oily solutions or suspensions), for administration byinhalation (for example as a finely divided powder or a liquid aerosol),for administration by insufflation (for example as a finely dividedpowder) or for parenteral administration (for example as a sterileaqueous or oily solution for intravenous, subcutaneous, intramuscular orintramuscular dosing or as a suppository for rectal dosing).

The compositions of the invention may be obtained by conventionalprocedures using conventional pharmaceutical excipients, well known inthe art. Thus, compositions intended for oral use may contain, forexample, one or more colouring, sweetening, flavouring and/orpreservative agents.

The compound of Formula (I) will normally be administered to awarm-blooded animal at a unit dose within the range 5-5000 mg/m² bodyarea of the animal, i.e. approximately 0.1-100 mg/kg, and this normallyprovides a therapeutically-effective dose. A unit dose form such as atablet or capsule will usually contain, for example 1-250 mg of activeingredient. Preferably a daily dose in the range of 1-50 mg/kg isemployed, for example 4-7 mg/kg twice daily. However the daily dose willnecessarily be varied depending upon the host treated, the particularroute of administration, and the severity of the illness being treated.Accordingly the practitioner who is treating any particular patient maydetermine the optimum dosage.

For example, a pharmaceutical composition of the present inventionsuitable for oral administration could comprise 1-200 mg/ml of acompound of Formula (I), or a pharmaceutically-acceptable salt thereof,(such as(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide)in 0.5% hydroxypropylmethylcellulose (HPMC).

In the context of the present specification, the term “therapy” alsoincludes “prophylaxis” unless there are specific indications to thecontrary. The terms “therapeutic” and “therapeutically” should beconstrued accordingly.

As used herein, the term “treatment” is intended to have its normaleveryday meaning of dealing with a disease in order to entirely orpartially relieve one, some or all of its symptoms, or to correct orcompensate for the underlying pathology.

As used herein, the term “prophylaxis” is intended to have its normaleveryday meaning and includes primary prophylaxis to prevent thedevelopment of the disease and secondary prophylaxis whereby the diseasehas already developed and the patient is temporarily or permanentlyprotected against exacerbation or worsening of the disease or thedevelopment of new symptoms associated with the disease.

As a result of their PKB inhibitory activity, the compounds of Formula(I) of the present invention are expected to be useful in the treatmentof diseases or medical conditions mediated alone or in part by PKBactivity, for example cancer. The types of cancers which may besusceptible to treatment using compounds of Formula (I) of the presentinvention include, but are not limited to, ovarian cancer, cervicalcancer, colorectal cancer, breast cancer, pancreatic cancer, glioma,glioblastoma, melanoma, prostate cancer, leukaemia, lymphoma,Non-Hodgkins lymphoma, gastric cancer, lung cancer, hepatocellularcancer, gastric cancer, gastrointestinal stromal tumour (GIST), glioma,thyroid cancer, bile duct cancer, endometrial cancer, renal cancer,anaplastic large cell lymphoma, acute myeloid leukaemia (AML), multiplemyeloma, melanoma and mesothelioma. Breast cancer, and more specificallyluminal breast cancer, may be particularly susceptible to treatmentusing compounds of the present invention.

It is envisaged that for the methods of treatment of cancer mentionedherein, the compound of Formula (I) will be administered to a mammal,more particularly a human being. Similarly, for the uses of a compoundof Formula (I) for the treatment of cancer mentioned herein, it isenvisaged that the compound of Formula (I) will be administered to amammal, more particularly a human being.

According to a another aspect of the invention, there is thereforeprovided a compound of Formula (I) as defined hereinbefore, or apharmaceutically acceptable salt thereof, for use as a medicament.

According to a further aspect of the invention, there is provided acompound of Formula (I) as defined hereinbefore, or a pharmaceuticallyacceptable salt thereof for use in the treatment of a disease mediatedthrough PKB. In one embodiment of the invention, said disease mediatedthrough PKB is cancer. In a further embodiment of the invention, saidcancer is selected from ovarian cancer, cervical cancer, colorectalcancer, breast cancer, pancreatic cancer, glioma, glioblastoma,melanoma, prostate cancer, leukaemia, lymphoma, Non-Hodgkins lymphoma,gastric cancer, lung cancer, hepatocellular cancer, gastric cancer,gastrointestinal stromal tumour (GIST), glioma, thyroid cancer, bileduct cancer, endometrial cancer, renal cancer, anaplastic large celllymphoma, acute myeloid leukaemia (AML), multiple myeloma, melanoma andmesothelioma. In one embodiment of the invention, said cancer isselected from breast cancer, Non-Hodgkins lymphoma, pancreatic cancer,hepatocellular cancer, gastric cancer, prostate cancer and lung cancer.In one particular embodiment, said cancer is breast cancer, moreparticularly luminal breast cancer.

According to a further aspect of the invention, there is provided theuse of a compound of Formula (I) as defined hereinbefore, or apharmaceutically acceptable salt thereof for the preparation of amedicament for the treatment of a disease mediated through PKB. In oneembodiment of the invention, said disease mediated through PKB iscancer. In a further embodiment of the invention, said cancer isselected from ovarian cancer, cervical cancer, colorectal cancer, breastcancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostatecancer, leukaemia, lymphoma, Non-Hodgkins lymphoma, gastric cancer, lungcancer, hepatocellular cancer, gastric cancer, gastrointestinal stromaltumour (GIST), glioma, thyroid cancer, bile duct cancer, endometrialcancer, renal cancer, anaplastic large cell lymphoma, acute myeloidleukaemia (AML), multiple myeloma, melanoma and mesothelioma. In oneembodiment of the invention, said cancer is selected from breast cancer,Non-Hodgkins lymphoma, pancreatic cancer, hepatocellular cancer, gastriccancer, prostate cancer and lung cancer. In one particular embodiment,said cancer is breast cancer, more particularly luminal breast cancer.

According to a further aspect of the invention, there is provided theuse of a compound of Formula (I) as defined hereinbefore, or apharmaceutically acceptable salt thereof, for the preparation of amedicament for the treatment of cancer. In one embodiment of theinvention, said cancer is selected from ovarian cancer, cervical cancer,colorectal cancer, breast cancer, pancreatic cancer, glioma,glioblastoma, melanoma, prostate cancer, leukaemia, lymphoma,Non-Hodgkins lymphoma, gastric cancer, lung cancer, hepatocellularcancer, gastric cancer, gastrointestinal stromal tumour (GIST), glioma,thyroid cancer, bile duct cancer, endometrial cancer, renal cancer,anaplastic large cell lymphoma, acute myeloid leukaemia (AML), multiplemyeloma, melanoma and mesothelioma. In one embodiment of the invention,said cancer is selected from breast cancer, Non-Hodgkins lymphoma,pancreatic cancer, hepatocellular cancer, gastric cancer, prostatecancer and lung cancer. In one particular embodiment, said cancer isbreast cancer, more particularly luminal breast cancer.

According to a further aspect of the invention, there is provided amethod of using a compound of Formula (I) as defined hereinbefore, or apharmaceutically acceptable salt thereof, for the treatment of cancer.In one embodiment of the invention, said cancer is selected from ovariancancer, cervical cancer, colorectal cancer, breast cancer, pancreaticcancer, glioma, glioblastoma, melanoma, prostate cancer, leukaemia,lymphoma, Non-Hodgkins lymphoma, gastric cancer, lung cancer,hepatocellular cancer, gastric cancer, gastrointestinal stromal tumour(GIST), glioma, thyroid cancer, bile duct cancer, endometrial cancer,renal cancer, anaplastic large cell lymphoma, acute myeloid leukaemia(AML), multiple myeloma, melanoma and mesothelioma. In one embodiment ofthe invention, said cancer is selected from breast cancer, Non-Hodgkinslymphoma, pancreatic cancer, hepatocellular cancer, gastric cancer,prostate cancer and lung cancer. In one particular embodiment, saidcancer is breast cancer, more particularly luminal breast cancer.

According to a further aspect of the invention, there is provided amethod of treating a human suffering from a disease in which inhibitionof PKB is beneficial, comprising the steps of administering to a personin need thereof of a therapeutically effective amount of a compound ofFormula (I) as defined hereinbefore, or a pharmaceutically acceptablesalt thereof. In one embodiment of the invention, the disease in whichinhibition of PKB is beneficial is cancer. In a further embodiment ofthe invention, said cancer is selected from ovarian cancer, cervicalcancer, colorectal cancer, breast cancer, pancreatic cancer, glioma,glioblastoma, melanoma, prostate cancer, leukaemia, lymphoma,Non-Hodgkins lymphoma, gastric cancer, lung cancer, hepatocellularcancer, gastric cancer, gastrointestinal stromal tumour (GIST), glioma,thyroid cancer, bile duct cancer, endometrial cancer, renal cancer,anaplastic large cell lymphoma, acute myeloid leukaemia (AML), multiplemyeloma, melanoma and mesothelioma. In one embodiment of the invention,said cancer is selected from breast cancer, Non-Hodgkins lymphoma,pancreatic cancer, hepatocellular cancer, gastric cancer, prostatecancer and lung cancer. In one particular embodiment, said cancer isbreast cancer, more particularly luminal breast cancer.

The cancer treatment defined hereinbefore may be applied as a soletherapy or may involve, in addition to the compound of the invention,conventional surgery or radiotherapy or chemotherapy. Such chemotherapymay include one or more of the following categories of anti-tumouragents:—

-   -   (i) other antiproliferative/antineoplastic drugs and        combinations thereof, as used in medical oncology, such as        alkylating agents (for example cis-platin, oxaliplatin,        carboplatin, cyclophosphamide, nitrogen mustard, melphalan,        chlorambucil, busulphan, temozolamide and nitrosoureas);        antimetabolites (for example gemcitabine and antifolates such as        fluoropyrimidines like 5-fluorouracil and tegafur, raltitrexed,        methotrexate, cytosine arabinoside, and hydroxyurea); antitumour        antibiotics (for example anthracyclines like adriamycin,        bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin,        mitomycin-C, dactinomycin and mithramycin); antimitotic agents        (for example vinca alkaloids like vincristine, vinblastine,        vindesine and vinorelbine and taxoids like taxol and taxotere        and polokinase inhibitors); and topoisomerase inhibitors (for        example epipodophyllotoxins like etoposide and teniposide,        amsacrine, topotecan and camptothecin);    -   (ii) cytostatic agents such as antioestrogens (for example        tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and        iodoxyfene), antiandrogens (for example bicalutamide, flutamide,        nilutamide and cyproterone acetate), LHRH antagonists or LHRH        agonists (for example goserelin, leuprorelin and buserelin),        progestogens (for example megestrol acetate), aromatase        inhibitors (for example as anastrozole, letrozole, vorazole and        exemestane) and inhibitors of 5α-reductase such as finasteride;    -   (iii) anti-invasion agents (for example c-Src kinase family        inhibitors like        4-(6-chloro-2,3-methylenedioxyanilino)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5-tetrahydropyran-4-yloxyquinazoline        (AZD0530; International Patent Application WO 01/94341) and        N-(2-chloro-6-methylphenyl)-2-{6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-ylamino}thiazole-5-carboxamide        (dasatinib, BMS-354825; J. Med. Chem., 2004, 47, 6658-6661), and        metalloproteinase inhibitors like marimastat, inhibitors of        urokinase plasminogen activator receptor function or antibodies        to Heparanase);    -   (iv) inhibitors of growth factor function: for example such        inhibitors include growth factor antibodies and growth factor        receptor antibodies (for example the anti-erbB2 antibody        trastuzumab [Herceptin™], the anti-EGFR antibody panitumumab,        the anti-erbB1 antibody cetuximab [Erbitux, C225] and any growth        factor or growth factor receptor antibodies disclosed by Stern        et al. Critical reviews in oncology/haematology, 2005, Vol. 54,        pp 11-29); such inhibitors also include tyrosine kinase        inhibitors, for example inhibitors of the epidermal growth        factor family (for example EGFR family tyrosine kinase        inhibitors such as        N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine        (gefitinib, ZD1839),        N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine        (erlotinib, OSI-774) and        6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazolin-4-amine        (CI 1033), erbB2 tyrosine kinase inhibitors such as lapatinib,        inhibitors of the hepatocyte growth factor family, inhibitors of        the platelet-derived growth factor family such as imatinib,        inhibitors of serine/threonine kinases (for example Ras/Raf        signalling inhibitors such as farnesyl transferase inhibitors,        for example sorafenib (BAY 43-9006)), inhibitors of cell        signalling through MEK and/or AKT kinases, inhibitors of the        hepatocyte growth factor family, c-kit inhibitors, abl kinase        inhibitors, IGF receptor (insulin-like growth factor) kinase        inhibitors; aurora kinase inhibitors (for example AZD1152,        PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528 AND        AX39459) and cyclin dependent kinase inhibitors such as CDK2        and/or CDK4 inhibitors;    -   (v) antiangiogenic agents such as those which inhibit the        effects of vascular endothelial growth factor, [for example the        anti-vascular endothelial cell growth factor antibody        bevacizumab (Avastin™) and VEGF receptor tyrosine kinase        inhibitors such as        4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline        (ZD6474; Example 2 within WO 01/32651),        4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-1-ylpropoxy)quinazoline        (AZD2171; Example 240 within WO 00/47212), vatalanib (PTK787; WO        98/35985) and SU11248 (sunitinib; WO 01/60814), compounds such        as those disclosed in International Patent Applications        WO97/22596, WO 97/30035, WO 97/32856 and WO 98/13354 and        compounds that work by other mechanisms (for example linomide,        inhibitors of integrin αvβ3 function and angiostatin)];    -   (vi) vascular damaging agents such as Combretastatin A4 and        compounds disclosed in International Patent Applications WO        99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 and        WO 02/08213;    -   (vii) antisense therapies, for example those which are directed        to the targets listed above, such as ISIS 2503, an anti-ras        antisense;    -   (viii) gene therapy approaches, including for example approaches        to replace aberrant genes such as aberrant p53 or aberrant BRCA1        or BRCA2, GDEPT (gene-directed enzyme pro-drug therapy)        approaches such as those using cytosine deaminase, thymidine        kinase or a bacterial nitroreductase enzyme and approaches to        increase patient tolerance to chemotherapy or radiotherapy such        as multi-drug resistance gene therapy; and    -   (ix) immunotherapy approaches, including for example ex-vivo and        in-vivo approaches to increase the immunogenicity of patient        tumour cells, such as transfection with cytokines such as        interleukin 2, interleukin 4 or granulocyte-macrophage colony        stimulating factor, approaches to decrease T-cell anergy,        approaches using transfected immune cells such as        cytokine-transfected dendritic cells, approaches using        cytokine-transfected tumour cell lines and approaches using        anti-idiotypic antibodies.

A compound of the invention, or a salt thereof, may be prepared by anyprocess known to be applicable to the preparation of such compounds orstructurally related compounds. Functional groups may be protected anddeprotected using conventional methods. For examples of protectinggroups such as amino and carboxylic acid protecting groups (as well asmeans of formation and eventual deprotection), see T. W. Greene and P.G. M. Wuts, “Protective Groups in Organic Synthesis”, Second Edition,John Wiley & Sons, New York, 1991.

Certain processes for the synthesis of compounds of Formula (I) areprovided as a further feature of the invention. Thus, according to afurther aspect of the invention there is provided a process for thepreparation of a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof, which comprises a process (a), (b), (c) or (d)(wherein the variables are as defined hereinbefore for compounds ofFormula (I) unless otherwise defined):

-   -   (a) reaction of an acid of Formula (II) with an amine of Formula        (III):

-   -   wherein P¹ represents a suitable protecting group, for example        tert-butoxycarbonyl;    -   (b) reaction of a carboxamide of Formula (IV) with a bicyclic        heterocycle of Formula (V):

-   -   wherein L₁ represents a suitable leaving group, for example        chlorine;    -   (c) when n is 1, hydrogenation of a compound of Formula (VI): or

-   -   (d) when R¹ represents aminomethyl, hydrogenation of a compound        of Formula (VII):

-   -   wherein P¹ represents a suitable protecting group, for example        tert-butoxycarbonyl;    -   and thereafter, if necessary:    -   (i) converting a compound of Formula (I) into another compound        of Formula (I);    -   (ii) removing any protecting groups; and/or    -   (iii) forming a pharmaceutically acceptable salt thereof.

Examples of conversions of a compound of Formula (I) into anothercompound of Formula (I), are well known to those skilled in the art, andinclude functional group interconversions such as hydrolysis,hydrogenation, hydrogenolysis, oxidation or reduction, and/or furtherfunctionalisation by standard reactions such as amide or metal-catalysedcoupling, or nucleophilic displacement reactions.

Specific reaction conditions for processes (a), (b), (c) and (d) aboveare as follows: Process (a)—acids of Formula (II) and amines of Formula(III) may be reacted together in the presence of a suitable couplingreagent, for exampleO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU), and a suitable base, for exampleN,N′-diisopropylethylamine (DIPEA), in a suitable solvent, for exampledimethylacetamide (DMA), and at a suitable temperature, for example 50to 70° C., more suitably about 60° C.;

-   -   Process (b)—carboxamides of Formula (IV) and heterocycles of        Formula (V) may be reacted together in the presence of a        suitable base, for example N,N′-diisopropylethylamine (DIPEA),        in a suitable solvent, for example butan-1-ol, and at a suitable        temperature, for example 50 to 70° C., more suitably about 60°        C.; Processes (c) and (d)—compounds of Formula (VI) or (VII)        dissolved in a suitable solvent, for example ethanol, may be        hydrogenated under an atmosphere of hydrogen in the presence of        a suitable catalyst, for example Raney™ nickel, and a suitable        base, for example ammonium hydroxide.

Compounds of Formula (II) may be prepared according to Scheme 1:

wherein P¹ is a suitable protecting group, for exampletert-butoxycarbonyl, L¹ is a suitable leaving group, for examplechlorine, and all other variables are as defined hereinbefore.

Compounds of Formula (IV) may be prepared according to Scheme 2:

wherein P¹ and P² are suitable protecting groups, for exampletert-butoxycarbonyl, and all other variables are as definedhereinbefore.

Compounds of Formula (VI) may be prepared according to Scheme 3:

wherein P¹ is a suitable protecting group, for exampletert-butoxycarbonyl, and all other variables are as definedhereinbefore.

Compounds of Formula (VII) may be prepared according to Scheme 4:

wherein P¹ represents a suitable protecting group, for exampletert-butoxycarbonyl, and all other variables are as definedhereinbefore.

Compounds of Formulas (III), (V), (VIII) and (IX) are commerciallyavailable, known in the literature, prepared by standard processes knownin the art, or may be prepared according to the processes describedherein.

The following examples are for illustration purposes and are notintended to limit the scope of this application. Each exemplifiedcompound represents a particular and independent aspect of theinvention. All starting materials are commercially available compounds,or they are known in the literature, or they are prepared by standardprocesses known in the art.

Generally, with respect to the following Examples:

-   -   (i) temperatures are given in degrees Celsius (° C.); operations        were carried out at room or ambient temperature, that is, at a        temperature in the range of 18 to 25° C.;    -   (ii) organic solutions were dried over anhydrous magnesium        sulfate or anhydrous sodium sulfate; evaporation of solvent was        carried out using a rotary evaporator under reduced pressure        (600 to 4000 Pascals; 4.5 to 30 mmHg) with a bath temperature of        up to 60° C.;    -   (iii) chromatography means flash chromatography on silica gel;        thin layer chromatography (TLC) was carried out on silica gel        plates;    -   (iv) in general, the course of reactions was followed by TLC        and/or analytical LC-MS, and reaction times where given are for        illustration only.    -   (v) final products had satisfactory proton nuclear magnetic        resonance (NMR) spectra and/or mass spectral data;    -   (vi) yields are given for illustration only and are not        necessarily those which can be obtained by diligent process        development; preparations were repeated if more material was        required;    -   (vii) when given, NMR data is in the form of delta values for        major diagnostic protons, given in parts per million (ppm)        relative to tetramethylsilane (TMS) as an internal standard,        determined at 500 MHz using perdeuterio dimethyl sulfoxide        (DMSO-d6) as solvent unless otherwise indicated; the following        abbreviations have been used: s, singlet; d, doublet; t,        triplet; q, quartet; m, multiplet; bs, broad singlet;    -   (viii) chemical symbols have their usual meanings; SI units and        symbols are used;    -   (ix) Mass spectra (MS) and LC-MS data were generated on an LC-MS        system where the HPLC component comprised generally either an        Agilent 1100, Waters Alliance HT (2790 & 2795) equipment or an        HP1100 pump and Diode Array with CTC autosampler and was run on        a Phenomenex Gemini C18 5 μm, 50×2 mm column (or similar)        eluting with either acidic eluent (for example, using a gradient        between 0-95% water/acetonitrile with 5% of a 1% formic acid in        50:50 water:acetonitrile (v/v) mixture), or basic eluent (for        example, using a gradient between 0-95% water/acetonitrile with        5% of a 0.1% 880 Ammonia in acetonitrile mixture); and the MS        component comprised generally a Waters ZQ mass spectrometer        scanning over an appropriate mass range. Chromatograms for        Electrospray (ESI) positive and negative Base Peak Intensity,        and UV Total Absorption Chromatogram from 220-300 nm, are        generated and values for m/z are given; generally, only ions        which indicate the parent mass are reported and unless otherwise        stated the value quoted is the (M+H)+ for positive ion mode and        (M−H)− for negative ion mode;    -   (x) unless stated otherwise compounds containing an        asymmetrically substituted carbon and/or sulfur atom have not        been resolved;    -   (xi) any microwave reactions were carried out in either a        Biotage Optimizer EXP, or a CEM Explorer microwave;    -   (xii) preparative high performance liquid chromatography (HPLC)        was performed on a Gilson instrument using the following        conditions:    -   Column: C₁₈ reversed-phase silica, for example, Waters        ‘Xbridge’, 5 μm silica, 19×100 mm, or 30×100 mm, using        decreasingly polar solvent mixtures as eluent (decreasing ratio        of Solvent A to Solvent B)    -   Solvent A: Water with 1% ammonium hydroxide    -   Solvent B: Acetonitrile    -   Flow rate: 28 ml/min or 61 ml/min    -   Gradient: Tailored to suit each compound—generally 7-10 min in        length    -   Wavelength: 254 nm

Abbreviations

-   -   Boc Tert-butoxycarbonyl    -   CAS™ Chemical Abstracts Service    -   DCM dichloromethane    -   DIPEA N,N′-diisopropylethylamine    -   DEA diethylamine    -   DMA dimethylacetamide    -   DMF dimethylformamide    -   HATU O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium        hexafluorophosphate    -   LCMS liquid chromatography mass spectroscopy    -   LDA lithium diisopropylamide    -   MPLC medium pressure liquid chromatography    -   NMP N-methylpyrrolidinone    -   OBD optimum bed density    -   PTFE polytetrafluoroethylene    -   SCX strong cation exchange    -   SFC supercritical flow chromatography    -   TBME t-butyl methyl ether    -   TEA triethylamine    -   TFA trifluoroacetic acid    -   THF tetrahydrofuran

EXAMPLE 14-amino-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

4-(tert-Butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (362 mg), 1-(4-chlorophenyl)ethanamine (172 mg),N-(3-dimethylaminopropyl)-3-ethylcarbodiimide (231 mg) and1-hydroxybenzotriazole (163 mg) were stirred together in DMF (2 mL)under nitrogen for 16 hours. The reaction mixture was partitionedbetween EtOAc (20 mL) and brine (4×20 mL). The organics were combined,dried over MgSO4 and evaporated in vacuo. The resultant white solid wasdissolved in 1,4-dioxane (5 mL) and a 4M solution of HCl in 1,4-dioxane(5 mL) was added. The resulting mixture was stirred for 16 hours, thendiluted with diethyl ether (50 mL). The crude product was isolated byfiltration as the HCl salt which was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 7M ammonia/MeOH and pure fractions were evaporated todryness. This material was purified by preparative LCMS usingdecreasingly polar mixtures of water (containing 1% NH₃) and MeCN aseluents. Fractions containing the desired compound were evaporated todryness to afford4-amino-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideas a white solid (168 mg, 42%). 1H NMR (d6-dmso, 400 MHz) 1.33-1.49 (m,5H), 1.84-2.04 (m, 2H), 2.12-2.22 (br s, 2H), 3.54 (t, 2H), 4.39 (t,2H), 4.81-4.92 (m, 1H), 6.55-6.59 (m, 1H), 7.13-7.18 (m, 1H), 7.31-7.39(m, 4H), 8.12 (s, 1H), 8.30 (d, 1H), 11.62 (s, 1H).

MS m/e MH+ 399.

EXAMPLE 2(S)-4-amino-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (0.418 g) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (0.361 g), (5)-1-(4-chlorophenyl)ethanamine (0.140mL) and DIPEA (0.524 mL) in DMA (10 mL) at 25° C. under nitrogen. Theresulting solution was stirred at 60° C. for 4 hours. The crude productwas purified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using 7M NH₃/MeOH and purefractions were evaporated to dryness. This crude material was thentreated with a 20% solution of TFA in DCM (10 mL) and stirred at roomtemperature. The crude product was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 7M ammonia/MeOH and pure fractions were evaporated todryness. This material was purified by preparative LCMS usingdecreasingly polar mixtures of water (containing 1% NH₃) and MeCN aseluents. Fractions containing the desired compound were evaporated todryness to afford(S)-4-amino-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideas a white solid (0.281 g, 70.4%). 1H NMR (400.13 MHz, DMSO-d6) δ 1.37(3H, d), 1.42-1.45 (2H, m), 1.88-2.01 (2H, m), 2.27 (2H, s), 3.49-3.59(2H, m), 4.34-4.44 (2H, m), 4.83-4.90 (1H, m), 6.57-6.58 (1H, m),7.14-7.16 (1H, m), 7.32-7.38 (4H, m), 8.12 (1H, s), 8.30 (1H, d), 11.62(1H, s). MS m/e MH⁺399.

EXAMPLE 34-amino-N-(1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (0.209 g) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (0.181 g), 1-(4-chlorophenyl)propan-1-amine (0.085g) and DIPEA (0.262 mL) in DMA (10 mL) at 25° C. under nitrogen. Theresulting solution was stirred at 60° C. for 4 hours. The crude productwas purified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using 7N ammonia/MeOH andpure fractions were evaporated to dryness. This crude material was thentreated with a 20% solution of TFA in DCM (10 mL) and stirred at roomtemperature for 2 hours. The crude product was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 7N ammonia/MeOH and pure fractions were evaporated todryness. This material was purified by preparative LCMS usingdecreasingly polar mixtures of water (containing 1% ammonia) and MeCN aseluents. Fractions containing the desired compound were evaporated todryness to afford4-amino-N-(1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideas a white solid (0.138 g, 66.8%).

¹H NMR (400.13 MHz, DMSO-d6) δ 0.87 (3H, t), 1.42-1.55 (2H, m),1.72-1.79 (2H, m), 1.91-2.05 (2H, m), 2.21 (2H, s), 3.54-3.62 (2H, m),4.38-4.45 (2H, m), 4.65-4.70 (1H, m), 6.61 (1H, dd), 7.18 (1H, dd),7.32-7.37 (4H, m), 8.31 (1H, d), 8.12 (1H, s). MS m/e MH⁺413.

EXAMPLES 3A AND 3B(S)-4-amino-N-(1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideand(R)-4-amino-N-(1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Racemic4-amino-N-(1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(Example 3) was purified by preparative chiral-HPLC. The fractionscontaining the desired compound were evaporated to dryness to affordIsomer 1 (first to elute, 41 mg) as a white solid, and Isomer 2 (secondto elute, 41 mg) as a white solid. Analytical data was identical to theoriginal sample. Chiral analytical HPLC analysis (using a 20 μmChiralpak AS (250 mm×4.6 mm) column, with an eluent mixture ofiso-hexane/(EtOH/MeOH 50/50)/TEA 90/10/0.1, 1 mL/min at 25° C.,injecting 10 μl of a 1 mg/mL solution in EtOH) showed each enantiomer tobe distinct from each other and enantiomerically pure (e.e. =100%).

EXAMPLE 44-amino-N-((4-chlorophenyl)(cyclopropyl)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (0.228 g) was added to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (0.181 g),(4-chlorophenyl)(cyclopropyl)methanamine (Intermediate 3) (0.091 g) andDIPEA (0.261 mL) in DMA (5 mL) at 25° C. The resulting solution wasstirred at 50° C. for 1 hour. The crude product was purified by ionexchange chromatography, using an SCX column. The desired product waseluted from the column using 7N ammonia/MeOH and pure fractions wereevaporated to dryness. This crude material was suspended indichloromethane (25 mL), and TFA (5 mL) was added. The reaction mixturewas stirred for 1 hour, then the crude product was purified by ionexchange chromatography, using an SCX column. The desired product waseluted from the column using 7N ammonia/MeOH and pure fractions wereevaporated to dryness to afford the crude material as a white solid.This material was triturated under cold methanol to give the pureproduct,4-amino-N-((4-chlorophenyl)(cyclopropyl)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideas a white solid (0.167 g, 79%).

¹H NMR (400.13 MHz, DMSO-d6) δ 0.27-0.37 (2H, m), 0.48-0.52 (2H, m),1.18-1.24 (1H, m), 1.40-1.48 (2H, m), 1.88-2.02 (2H, m), 2.20 (2H, s),3.50-3.59 (2H, m), 4.15 (1H, t), 4.36-4.42 (2H, m), 6.57-6.58 (1H, m),7.14-7.16 (1H, m), 7.35-7.40 (4H, m), 8.12 (1H, s), 8.47 (1H, d), 11.62(1H, s).

MS m/e MH⁺425.

EXAMPLE 54-amino-N-(2-amino-1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Trifluoroacetic acid (3 mL) was added to tert-butyl4-(2-amino-1-(4-chlorophenyl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 6) (0.514 g) in DCM (100 mL) at 25° C. The resultingsolution was stirred at 25° C. for 3 hours. The crude product waspurified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using 7M NH₃/MeOH and purefractions were evaporated to dryness. The crude product was purified bypreparative HPLC (Waters XBridge Prep C18 OBD column, 5 μl silica, 19 mmdiameter, 100 mm length), using decreasingly polar mixtures of water(containing 1% NH3) and MeCN as eluents. Fractions containing thedesired compound were evaporated to dryness. This material was thenfurther purified by flash silica chromatography, elution gradient 0 to10% methanolic ammonia (71V) in DCM. Pure fractions were evaporated todryness to afford4-amino-N-(2-amino-1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideas a colourless gum (0.047 g, 11.4%). 1H NMR (400.13 MHz, DMSO-d6) δ1.41-1.51 (2H, m), 1.86-1.94 (2H, m), 1.97-2.05 (2H, m), 2.77-2.86 (2H,m), 3.18 (2H, s), 3.51-3.59 (2H, m), 4.35-4.44 (2H, m), 4.70 (1H, t),6.58 (1H, d), 7.15 (1H, d), 7.30-7.32 (2H, m), 7.35-7.37 (2H, m), 8.12(1H, s), 8.41 (1H, s), 11.62 (1H, s).

MS m/e MH⁺414.

EXAMPLE 6(S)-4-(aminomethyl)-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Trifluoroacetic acid (2 mL, 25.96 mmol) was added to (S)-tert-butyl(4-(1-(4-chlorophenyl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-yl)methylcarbamate(Intermediate 13) (0.257 g, 0.5 mmol) in DCM (20 mL) at 25° C. Theresulting solution was stirred at 25° C. for 1 hour. The crude productwas purified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using 7M NH₃/MeOH and purefractions were evaporated to dryness. The crude product was purified byflash silica chromatography, elution gradient 0 to 10% 7N ammonia/MeOHin DCM. Pure fractions were evaporated to dryness to afford(S)-4-(aminomethyl)-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(0.095 g, 46.0%) as a white solid.

1H NMR (400.13 MHz, DMSO-d6) δ 1.39 (3H, d), 1.43-1.50 (4H, m),2.05-2.14 (2H, m), 2.69 (2H, s), 3.37-3.47 (2H, m), 4.20-4.25 (2H, m),4.96-5.04 (1H, t), 6.55 (1H, d), 7.15 (1H, d), 7.37 (4H, s), 8.11 (1H,s), 8.49 (1H, d), 11.61 (1H, s).

MS m/e MH⁺ 413.

EXAMPLE 74-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Trifluoroacetic acid (2 ml, 25.96 mmol) was added to tert-butyl4-(1-(4-chlorophenyl)-4-hydroxybutylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 17) (140 mg, 0.26 mmol) at 20° C. and the resultingsolution stirred for 1 hour. The solution was then diluted withmethanol, applied to a 10 g SCX column and eluted with methanol followedby 2N NH₃/MeOH. Fractions containing product were combined, concentratedby evaporation and purified by preparative HPLC (Waters XBridge Prep C18OBD column, 5 μm silica, 19 mm diameter, 100 mm length), usingdecreasingly polar mixtures of water (containing 1% NH₃) and MeCN aseluents. Fractions containing the desired compound were evaporated todryness to afford4-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(58.0 mg, 50.8%) as a colourless solid.

1H NMR (399.902 MHz, DMSO) δ 1.28-1.51 (4H, m), 1.69-1.80 (2H, m),1.90-2.03 (2H, m), 3.37-3.41 (2H, m), 3.50-3.58 (2H, m), 4.37-4.43 (3H,m), 4.71-4.76 (1H, m), 6.59 (1H, m), 7.16 (1H, m), 7.36 (4H, m), 8.13(1H, s), 8.33 (1H, d), 11.64 (1H, s).

MS m/e MH⁺443.

EXAMPLES 7A AND 7B(S)-4-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideand(R)-4-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

DIPEA (2.85 mL, 16.0 mmol) was added to4-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)piperidine-4-carboxamide(Intermediate 72) (1.04 g, 3.19 mmol) and4-chloro-7H-pyrrolo[2,3-d]pyrimidine (0.490 g, 3.19 mmol) in ethanol(15.96 mL) at 25° C. The resulting solution was stirred at 65° C.overnight. The crude product was analysed by LCMS and evaporated todryness. The crude solid was then purified by ion exchangechromatography, using an SCX-2 column. The product was eluted from thecolumn using 20% 7N ammonia in methanol/DCM. The crude mixture was thenre-purified by flash silica chromatography (eluent 0-10% 7N ammonia/MeOHin DCM) and pure fractions evaporated to dryness to afford4-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(63.9%) (racemate) as a fine white solid. The racemate was chirallyresolved by supercritical flow chromatography (SFC) to afford the pureenantiomers in yields of 272 mg (19%) and 245 mg (17%) respectively. TheNMR spectra for both enantiomers were identical.

¹H NMR (400.13 MHz, DMSO) δ 1.38-1.42 (2H, m), 1.46-1.49 (2H, d), 1.74(2H, s), 1.92-2.03 (2H, m), 2.19 (2H, s), 3.55-3.58 (2H, d), 4.38 (1H,s), 4.41 (2H, s), 4.75-4.76 (1H, d), 6.59 (1H, s), 7.17 (1H, s), 7.36(4H, s), 8.15 (1H, s), 8.32-8.34 (1H, d), 11.65 (1H, s, exchange);

MS m/e MH⁺443; HPLC t_(R)=1.66 min.

EXAMPLE 84-amino-N-(1-(4-chlorophenyl)-2-hydroxyethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

tert-Butyl4-(1-(4-chlorophenyl)-2-hydroxyethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 18) (137 mg, 0.27 mmol) was treated with trifluoroaceticacid (2 mL). The solution was stirred for 1 hour at room temperature.The mixture was concentrated under reduced pressure. The crude productwas purified by ion exchange chromatography, using a SCX column. Theresidue was loaded onto the column in methanol and washed with methanol.The desired product was eluted from the column using 2M ammonia inmethanol and pure fractions were evaporated to dryness to afford4-amino-N-(1-(4-chlorophenyl)-2-hydroxyethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(111 mg, quant.) as a colourless crystalline solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.40-1.49 (2H, m), 1.85-2.09 (2H, m),3.48-3.69 (4H, m), 4.35-4.48 (2H, m), 4.72-4.81 (1H, m), 4.90-4.96 (1H,m), 6.58 (1H, br, s), 7.12-7.18 (1H, m), 7.30-7.40 (4H, m), 8.13 (1H,s), 8.45-8.53 (1H, m), 11.64 (1H, s) m/z (ESI+) (M+H)+=415; HPLCt_(R)=1.57 min.

EXAMPLE 9(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(E9)

HCl (4M in Dioxane) (3.00 mL, 12.00 mmol) was added to (S)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 22) (1.27 g, 2.40 mmol) in dichloromethane (20 mL). Theresulting suspension was stirred at 20° C. for 16 hours. The reactionmixture was filtered through a PTFE filtercup and the crude solid waspurified by preparative HPLC (Waters XTerra C18 column, 5 μm silica, 19mm diameter, 100 mm length), using decreasingly polar mixtures of water(containing 1% TFA) and MeCN as eluents. Fractions containing thedesired compound were purified by ion exchange chromatography, using anSCX column. The desired product was eluted from the column using 7MNH₃/MeOH and pure fractions were evaporated to dryness to afford(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(0.200 g, 19.4%) as a white solid. 1H NMR (399.9 MHz, DMSO-d6) δ 1.45(2H, d), 1.86 (1H, d), 1.90-1.93 (1H, m), 2.19 (2H, s), 3.38 (2H, q),3.51-3.58 (2H, m), 4.35-4.38 (2H, m), 4.53 (1H, t), 4.88 (1H, d), 6.58(1H, t), 7.16 (1H, t), 7.32-7.38 (4H, m), 8.12 (1H, s), 8.43 (1H, d),11.63 (1H, s), m/z (ESI+) (M+H)+=429; HPLC tR=1.46 min.

EXAMPLE 9 ALTERNATIVE ROUTE 1(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

N-Ethyldiisopropylamine (1.676 ml, 9.62 mmol) was added to(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide(Intermediate 49) (1 g, 3.21 mmol) and4-chloro-7H-pyrrolo[2,3-d]pyrimidine (0.493 g, 3.21 mmol) in butan-1-ol(15 ml). The resulting solution was stirred at 60° C. for 18 hours. Thereaction mixture was diluted with EtOAc (50 mL), and washed sequentiallywith water (25 mL) and saturated brine (25 mL). The organic layer wasdried over MgSO₄, filtered and evaporated to afford crude product. Thecrude product was purified by flash silica chromatography, elutiongradient 0 to 6% MeOH with ammonia in DCM. Pure fractions wereevaporated to dryness to afford(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(842 mg) as a white foam.(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamidewas stirred in ethyl acetate (7 mL) for 18 hours. The solid wascollected by filtration, washed with a small amount of ethyl acetate andvacuum oven dried at 55° C. for 18 hours to afford(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(0.585 g, 42.5%) as a white solid.

m/z (ES+) (M+H)+=429; HPLC tR=1.60 min.

1H NMR (400.13 MHz, DMSO-d6) δ 1.39-1.47 (2H, m), 1.80-2.02 (4H, m),2.17 (2H, s), 3.35-3.40 (2H, m), 3.50-3.59 (2H, m), 4.34-4.41 (2H, m),4.53 (1H, t), 4.88 (1H, d), 6.57 (1H, m), 7.14-7.16 (1H, m), 7.31-7.37(4H, m), 8.12 (1H, s), 8.42 (1H, d), 11.62 (1H, s)

EXAMPLE 9 ALTERNATIVE ROUTE 2(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

(S)-3-Amino-3-(4-chlorophenyl)propan-1-ol (Intermediate 47) (2.055 g,11.07 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (4 g, 11.07 mmol) and DIPEA (5.80 ml, 33.20 mmol)in DMA (40 ml). HATU (4.63 g, 12.18 mmol) was added and the resultingsolution was stirred at 20° C. for 24 hours. The reaction mixture wasevaporated to dryness then diluted with EtOAc (300 mL), and washedsequentially with water (50 mL) and saturated brine (50 mL). The organiclayer was dried over MgSO₄, filtered and evaporated to afford crudeproduct. The crude product was purified by flash silica chromatography,elution gradient 2 to 6% MeOH with ammonia in DCM. Pure fractions wereevaporated to dryness and triturated with dioxane (40 ml) to afford(S)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 22) (4.82 g, 82%) as a white solid. (S)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 22) (4.82 g, 82%) was suspended in dioxane (40.0 ml) and4M hydrogen chloride in dioxane (7.69 ml, 221.36 mmol) added. Thereaction was stirred at ambient temperature for 2 hours. The crudeproduct was purified by ion exchange chromatography, using an SCXcolumn. The desired product was eluted from the column using 3.5MNH₃/MeOH and pure fractions were evaporated to dryness. The crudeproduct was purified by preparative HPLC (Waters XBridge Prep C18 OBDcolumn, 5 μm silica, 19 mm diameter, 100 mm length), using decreasinglypolar mixtures of water (containing 1% NH₃) and MeCN as eluents.Fractions containing the desired compound were evaporated to dryness toafford(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(1.200 g, 25.3%) as a white solid.

m/z (ES+) (M+H)+=429; HPLC tR=1.67 min.

¹H NMR matches previous.

EXAMPLE 10N-(3-acetamido-1-(4-chlorophenyl)propyl)-4-amino-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Hydrogen chloride (4M) in 1,4-Dioxane (0.430 mL, 1.72 mmol) was added totert-butyl4-(3-acetamido-1-(4-chlorophenyl)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 28) (98 mg, 0.17 mmol) in DCM (4 mL) at 20° C. Theresulting suspension was stirred at 20° C. for 70 hours. The reactionmixture was evaporated to dryness. The crude product was purified bypreparative HPLC (Waters XTerra C18 column, 5μ silica, 19 mm diameter,100 mm length), using decreasingly polar mixtures of water (containing1% NH₃) and MeCN as eluents. Fractions containing the desired compoundwere evaporated to dryness to affordN-(3-acetamido-1-(4-chlorophenyl)propyl)-4-amino-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(15.00 mg, 18.5%) as a white dry film. 1H NMR (399.9 MHz, DMSO-d6) δ1.42-1.47 (2H, m), 1.79 (3H, s), 1.85 (1H, t), 1.89-1.93 (1H, m), 2.10(2H, s), 3.00 (2H, t), 3.55 (2H, d), 4.36-4.40 (2H, m), 4.80 (1H, d),6.58-6.60 (1H, m), 7.16 (1H, t), 7.34-7.39 (4H, m), 7.80 (1H, t), 8.13(1H, s), 8.39 (1H, s), 11.64 (1H, s) No visible NH2. m/z (ESI+)(M+H)+=470; HPLC tR=1.56 min.

EXAMPLE 114-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Hydrogen chloride (4M in 1,4-dioxane, 1.01 mL, 4.05 mmol) was added totert-butyl4-(1-(4-chlorophenyl)-3-(dimethylamino)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 34) (0.045 g, 0.08 mmol) in a mixture of DCM (5 mL) andmethanol (2 mL) at 22° C. The resulting solution was stirred at 22° C.for 16 hours. The mixture was evaporated and the residue was purified byion exchange chromatography, using an SCX column. The desired productwas eluted from the column using 2M NH₃/MeOH and pure fractions wereevaporated to dryness to afford4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(0.034 g, 92%) as a colourless gum.

1H NMR (399.902 MHz, CDCl₃) δ 1.57 (2H, m), 1.66 (2H, br.s), 1.81 (1H,m), 2.02 (1H, m), 2.18 (6H, s), 2.18-2.36 (4H, m), 3.67 (3H, m), 4.50(2H, m), 5.00 (1H, dt), 6.52 (1H, d), 7.05 (1H, d), 7.18 (2H, d), 7.29(2H, d), 8.33 (1H, s), 9.07 (1H, d), 9.61 (1H, s).

MS m/e MH⁺456

EXAMPLES 11A AND 11B(S)-4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideand(R)-4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Racemic4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(Example 11) (231 mg, 0.51 mmol) was chirally separated on a ChiralpakAD-H SFC (250 mm×20 mm) column, using supercritical fluidchromatography, elution solvent 7:3 CO₂/(EtOH+0.1% DEA). The appropriatefractions for the first eluted isomer were evaporated and the residuetriturated with diethyl ether to give4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(59 mg, 25%) as a white solid.

1H NMR (399.902 MHz, DMSO) δ 1.43 (2H, ddd), 1.83 (2H, dt), 1.86-2.01(2H, m), 2.11 (6H, s), 2.14 (2H, t), 3.56 (2H, ddd), 4.39 (2H, ddd),4.83 (1H, dt), 6.58 (1H, dd), 7.16 (1H, dd), 7.33 (2H, d), 7.37 (2H, d),8.13 (1H, s), 8.61 (1H, d), 11.63 (1H, s).

MS m/e MH⁺456.5.

The appropriate fractions for the second eluted isomer were evaporatedand the residue triturated with diethyl ether to give4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(43 mg, 19%) as a white solid.

1H NMR (399.902 MHz, DMSO) δ 1.43 (2H, ddd), 1.83 (2H, dt), 1.86-2.01(2H, m), 2.11 (6H, s), 2.14 (2H, t), 3.56 (2H, ddd), 4.39 (2H, ddd),4.83 (1H, q), 6.58 (1H, dd), 7.16 (1H, dd), 7.33 (2H, d), 7.37 (2H, d),8.13 (1H, s), 8.61 (1H, d), 11.63 (1H, s).

MS m/e MH⁺456.4, 458.4

EXAMPLE 124-(aminomethyl)-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (106 mg, 0.28 mmol) was added to4-((tert-butoxycarbonylamino)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 12) (100 mg, 0.27 mmol) and N,N-diisopropylethylamine(0.055 mL, 0.32 mmol) in NMP (5 mL) at 22° C. The resulting solution wasstirred at 50° C. for 10 minutes then cooled to ambient temperature.3-Amino-3-(4-chlorophenyl)propan-1-ol (Intermediate 29) (49.5 mg, 0.27mmol) was added as a solution in NMP (2 mL) and the mixture was stirredat 22° C. for 16 hours. 4M HCl in dioxane (1 mL) was added and themixture was stirred at 22° C. for a further 24 hours. The mixture wasconcentrated and the residue was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 30% (2M NH₃ in MeOH) in DCM and pure fractions wereevaporated to dryness to afford crude product. The crude product waspurified by preparative HPLC (Waters XBridge Prep C18 OBD column, 5μsilica, 19 mm diameter, 100 mm length), using decreasingly polarmixtures of water (containing 1% NH₃) and MeCN as eluents. Fractionscontaining the desired compound were evaporated to dryness to afford4-(aminomethyl)-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(0.095 g, 81%) as a white solid.

1H NMR (399.902 MHz, DMSO) δ 1.47 (2H, m), 1.80-1.96 (2H, m), 2.09 (2H,m), 2.69 (1H, s), 3.27-3.45 (4H, m), 4.26 (2H, ddd), 5.02 (1H, dd), 6.56(1H, d), 7.15 (1H, d), 7.33-7.39 (4H, m), 8.12 (1H, s), 8.44 (1H, d),11.63 (1H, s).

MS m/e MH⁺556

EXAMPLE 134-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

TFA (0.7 mL) was added to a suspension of tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 36) (175 mg, 0.33 mmol) in dichloromethane (7 mL) underargon. The resulting solution was stirred at 20° C. for 16 hours. Thesolvents were removed in vacuo and the reaction mixture was purified bypreparative HPLC using a Waters X-Bridge reverse-phase column (C-18, 5microns silica, 19 mm diameter, 100 mm length) with decreasingly polarmixtures of water (containing 0.2% ammonium carbonate) and acetonitrileas eluent. The pure fractions were evaporated to dryness to afford4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(99 mg, 69.8%) as a white powder.

1H NMR (500 MHz, DMSO-d6) δ 1.38-1.46 (2H, m), 1.81-1.91 (4H, m), 2.25(2H, br s), 3.48 (2H, m), 3.35-3.54 (2H, m), 4.35-4.41 (2H, m), 4.57(1H, t), 4.87 (1H, m), 6.58 (1H, d), 7.16 (1H, d), 7.31-7.37 (4H, m),8.11 (1H, s), 8.45 (1H, d), 11.65 (1H, s). m/z (ESI+) (M+H)+=429; HPLCtR=1.58 min.

EXAMPLE 144-amino-N-(3-amino-1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Trifluoroacetic acid (0.05 mL) was added to a stirred suspension oftert-butyl4-(3-amino-1-(4-chlorophenyl)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 38) (7.0 mg) in DCM (1 mL) under argon at 25° C. Theresulting suspension was stirred at 25° C. for 2 days. The reactionmixture was evaporated to dryness to afford the di-TFA salt of4-amino-N-(3-amino-1-(4-chlorophenyl)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideas a partially solid oil (13.0 mg).

¹H NMR (500 MHz, DMSO-d6) δ 1.86 (1H, d), 2.01 (2H, m), 2.08 (1H, m)2.41 (2H, m), 2.69 (2H, m), 2.85 (1H, m), 3.61 (2H, t), 4.63 (2H, t),5.02 (1H, q), 6.82 (1H, s), 7.36 (1H, t), 7.41 (2H, d), 7.49 (2H, d),7.93 (3H, s br), 8.32 (1H, s), 8.59 (3H, s br), 8.96 (1H, d).

MS m/e MH⁺428.

EXAMPLE 15(R)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

HCl (4M in dioxane) (0.378 mL, 1.51 mmol) was added to (R)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 42) (0.160 g, 0.30 mmol) in DCM (3 mL). The resultingsuspension was stirred at 20° C. for 3 hours. The reaction mixture wasevaporated. The crude product was purified by preparative HPLC (WatersXTerra C18 column, 5 μm silica, 19 mm diameter, 100 mm length), usingdecreasingly polar mixtures of water (containing 1% ammonia) and MeCN aseluents. Fractions containing the desired compound were evaporated todryness to afford(R)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(0.051 g, 39.3%) as a white solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.46 (2H, d), 1.86 (1H, d), 1.90-1.93 (1H,m), 2.10 (2H, m), 3.37 (1H, t), 3.55 (2H, d), 4.40 (2H, d), 4.53 (2H,m), 4.88 (1H, d), 6.58 (1H, t), 7.16 (1H, t), 7.32-7.38 (4H, m), 8.14(1H, d), 8.43 (1H, d), 11.63 (1H, s) no visible NH₂.

MS m/e MH⁺429; HPLC tR=1.46 min.

EXAMPLE 16(R)-4-amino-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (0.418 g, 1.10 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (0.361 g, 1 mmol),(R)-1-(4-chlorophenyl)ethanamine (0.156 g, 1.00 mmol) and DIPEA (0.524mL, 3.00 mmol) in DMA (10 mL) at 25° C. under nitrogen. The resultingsolution was stirred at 60° C. for 4 hours. The crude product waspurified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using 7N ammonia/MeOH andpure fractions were evaporated to dryness. The crude material wastreated with a 10% solution of TFA in DCM (5 mL) and stirred at roomtemperature. The crude product was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 7N ammonia/MeOH and pure fractions were evaporated todryness. The crude product was purified by preparative LCMS usingdecreasingly polar mixtures of water (containing 1% ammonia) and MeCN aseluents. Fractions containing the desired compound were evaporated todryness to afford(R)-4-amino-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(0.211 g, 52.9%) as a white solid.

¹H NMR (400 MHz, DMSO) δ 1.37 (3H, d), 1.39-1.48 (2H, m), 1.86-2.02 (2H,m), 2.19 (2H, s), 3.49-3.58 (2H, m), 4.34-4.43 (2H, m), 4.83-4.91 (1H,m), 6.56-6.59 (1H, m), 7.14-7.16 (1H, m), 7.32-7.38 (4H, m), 8.12 (1H,s), 8.30 (1H, d), 11.62 (1H, s).

MS m/e MH⁺399.

EXAMPLE 17(R)-4-(aminomethyl)-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Raney® nickel (293 mg, 1.71 mmol) was added to a solution of(R)—N-(1-(4-chlorophenyl)ethyl)-4-cyano-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(Intermediate 45) (466 mg, 1.14 mmol), in ethanol (30 mL). Ammoniumhydroxide (10 mL) was added. This mixture was first purged withnitrogen, then placed under a balloon of hydrogen and stirred for 36hours. The reaction mixture was filtered through celite and the solventevaporated to dryness. The crude product was purified by flash silicachromatography, elution gradient 0 to 10% 7N ammonia/MeOH in DCM. Purefractions were evaporated to dryness to afford(R)-4-(aminomethyl)-N-(1-(4-chlorophenyl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(276 mg, 58.6%) as a white solid.

¹H NMR (400 MHz, DMSO) δ 1.38 (3H, d), 1.41-1.51 (2H, m), 1.61 (2H, s),2.09 (2H, d), 2.68 (2H, s), 3.37-3.50 (2H, m), 4.18-4.28 (2H, m),4.95-5.04 (1H, m), 6.56 (1H, d), 7.15 (1H, d), 7.26-7.50 (4H, m), 8.11(1H, s), 8.52 (1H, d), 11.66 (1H, s).

MS m/e MH⁺413.

EXAMPLE 18(S)-4-amino-N-(1-(4-cyanophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

(S)-4-(1-amino-3-hydroxypropyl)benzonitrile (Intermediate 46) (195 mg,1.11 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (400 mg, 1.11 mmol) and DIPEA (0.580 mL, 3.32mmol) in DMA (5 mL).O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (463 mg, 1.22 mmol) was added and the resultingsolution was stirred at 20° C. for 24 hours. The reaction mixture wasevaporated to dryness then diluted with EtOAc (300 mL), and washedsequentially with water (50 mL) and saturated brine (50 mL). The organiclayer was dried over MgSO₄, filtered and evaporated to afford crudeproduct. The crude product was purified by flash silica chromatography,elution gradient 2 to 6% MeOH with ammonia in DCM. Pure fractions wereevaporated to dryness to afford (S)-tert-butyl4-(1-(4-cyanophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(267 mg, 46.4%) as a white solid. (S)-tert-butyl4-(1-(4-cyanophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(267 mg, 0.51 mmol) was suspended in dioxane (5.00 mL) and 4M hydrogenchloride in dioxane (0.769 mL, 22.14 mmol) added. The reaction wasstirred at ambient temperature for 2 hours. The crude product waspurified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using 3.5N ammonia/MeOH andpure fractions were evaporated to dryness. The product was then purifiedby preparative HPLC (Waters XBridge Prep C18 OBD column, 5μ silica, 21mm diameter, 100 mm length), using decreasingly polar mixtures of water(containing 1% ammonia) and MeCN as eluents. Fractions containing thedesired compound were evaporated to dryness to afford(S)-4-amino-N-(1-(4-cyanophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(70.0 mg, 15.1%) as a white solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 1.36-1.47 (2H, m), 1.80-2.01 (4H, m),2.17 (2H, s), 3.39 (2H, q), 3.52-3.59 (2H, m), 4.34-4.40 (2H, m), 4.58(1H, t), 4.94 (1H, s), 6.57 (1H, d), 7.15 (1H, d), 7.50 (2H, d),7.76-7.79 (2H, d), 8.12 (1H, s), 8.52 (1H, s), 11.61 (1H, s).

MS m/e MH⁺420.

EXAMPLE 19(S)-4-amino-1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N-(1-(4-chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide

DIPEA (0.670 mL, 3.85 mmol) was added to(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide(Intermediate 49) (400 mg, 1.28 mmol) and5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (Intermediate 50) (298 mg,1.28 mmol) in butan-1-ol (6 mL). The resulting solution was stirred at60° C. for 18 hours. The reaction mixture was diluted with EtOAc (50mL), and washed sequentially with water (25 mL) and saturated brine (25mL). The organic layer was dried over MgSO₄, filtered and evaporated toafford crude product. The crude product was purified by flash silicachromatography, elution gradient 0 to 6% MeOH with ammonia in DCM. Theproduct was then purified by preparative HPLC (Waters XBridge Prep C18OBD column, 5μ silica, 19 mm diameter, 100 mm length), usingdecreasingly polar mixtures of water (containing 1% ammonia) and MeCN aseluents. Fractions containing the desired compound were evaporated todryness and triturated with diethyl ether to afford(S)-4-amino-1-(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N-(1-(4-chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide(70.0 mg, 10.8%) as a white solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 1.41-1.50 (2H, m), 1.81-1.92 (2H, m),2.04-2.20 (4H, m), 3.37-3.44 (4H, m), 3.91 (2H, t), 4.54 (1H, t), 4.90(1H, m), 7.33-7.38 (4H, m), 7.50 (1H, s), 8.25 (1H, s), 8.47 (1H, d),12.18 (1H, s).

MS m/e MH⁺510.

EXAMPLE 20(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)piperidine-4-carboxamide

DIPEA (0.419 mL, 2.41 mmol) was added to(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide(Intermediate 49) (250 mg, 0.80 mmol) and4-chloro-1H-pyrazolo[3,4-d]pyrimidine (124 mg, 0.80 mmol) in butan-1-ol(5 mL). The resulting solution was stirred at 60° C. for 6 hours. Thereaction mixture was diluted with EtOAc (50 mL), and washed sequentiallywith water (25 mL) and saturated brine (25 mL). The organic layer wasdried over MgSO₄, filtered and evaporated to afford crude product. Thecrude product was purified by flash silica chromatography, elutiongradient 0 to 6% MeOH with ammonia in DCM. Pure fractions wereevaporated to dryness and triturated with diethyl ether to afford(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)piperidine-4-carboxamide(112 mg, 32.5%) as a white solid.

1H NMR (399.9 MHz, DMSO-d6) δ 1.49 (2H, t), 1.79-2.01 (4H, m), 3.39 (2H,m), 3.62 (2H, s), 4.40 (2H, s), 4.57 (1H, t), 4.88 (1H, m), 7.32-7.38(4H, m), 8.22 (1H, d), 8.29 (1H, s), 8.45 (1H, d), 13.52 (1H, s).

MS m/e MH⁺430.

EXAMPLE 21(S)-4-amino-N-(3-hydroxy-1-phenylpropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

(S)-3-Amino-3-phenylpropan-1-ol hydrochloride (260 mg, 1.38 mmol) wasadded in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 21) (500 mg, 1.38 mmol) and DIPEA (0.967 mL, 5.53mmol) in DMA (5 mL).O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (579 mg, 1.52 mmol) was added and the resultingsolution was stirred at 20° C. for 24 hours. The reaction mixture wasevaporated to dryness then diluted with EtOAc (300 mL), and washedsequentially with water (50 mL) and saturated brine (50 mL). The organiclayer was dried over MgSO₄, filtered and evaporated to afford crudeproduct. The crude product was purified by flash silica chromatography,elution gradient 2 to 6% MeOH with ammonia in DCM. Fractions wereevaporated to afford (S)-tert-butyl4-(3-hydroxy-1-phenylpropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(334 mg, 48.8%) as a white solid. The product (334 mg, 0.67 mmol) wassuspended in dioxane (5.00 mL) and 4M hydrogen chloride in dioxane(0.961 mL, 27.67 mmol) added. The reaction was stirred at ambienttemperature for 2 hours. The crude product was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 3.5N ammonia/MeOH and pure fractions were evaporated todryness. The resulting gum was triturated with EtOAc to give a solidwhich was collected by filtration and dried under vacuum to give(S)-4-amino-N-(3-hydroxy-1-phenylpropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(93 mg, 17.0%) as an off white solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 1.44 (2H, t), 1.82-2.03 (4H, m), 3.37(2H, t), 3.50-3.58 (2H, m), 4.40 (2H, t), 4.49 (1H, t), 4.90 (1H, d),6.58 (1H, s), 7.15 (1H, t), 7.21-7.23 (1H, m), 7.30-7.31 (4H, m), 8.12(1H, s), 8.41 (1H, d), 11.62 (1H, s).

MS m/e MH⁺395.

EXAMPLE 22(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(9H-purin-6-yl)piperidine-4-carboxamide

DIPEA (0.335 mL, 1.92 mmol) was added to(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide(Intermediate 49) (200 mg, 0.64 mmol) and 6-chloro-9H-purine (99 mg,0.64 mmol) in butan-1-ol (4 mL). The resulting solution was stirred at60° C. for 18 hours. The reaction mixture was diluted with EtOAc (50mL), and washed sequentially with water (25 mL) and saturated brine (25mL). The organic layer was dried over MgSO₄, filtered and evaporated toafford crude product. The crude product was purified by flash silicachromatography, elution gradient 0 to 6% MeOH with ammonia in DCM. Purefractions were evaporated to dryness to afford(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(9H-purin-6-yl)piperidine-4-carboxamide(141 mg, 51.1%) as a white solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 1.42 (2H, t), 1.78-2.00 (4H, m),3.34-3.41 (2H, m), 3.61 (2H, s), 4.53 (1H, t), 4.88 (1H, d), 5.02 (2H,s), 7.30-7.39 (4H, m), 8.08 (1H, s), 8.17-8.22 (1H, s), 8.42 (1H, d),13.02 (1H, s).

MS m/e MH⁺430.

EXAMPLE 23(S)-4-(aminomethyl)-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

(S)-3-Amino-3-(4-chlorophenyl)propan-1-ol (Intermediate 47) (247 mg,1.33 mmol) was added in one portion to4-((tert-butoxycarbonylamino)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 12) (500 mg, 1.33 mmol) and DIPEA (0.698 mL, 4.00mmol) in DMA (6 mL).O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (557 mg, 1.47 mmol) was added and the resultingsolution was stirred at 20° C. for 2 hours then 20° C. for 18 hours. Thereaction mixture was diluted with EtOAc (50 mL), and washed sequentiallywith water (20 mL) and saturated brine (20 mL). The organic layer wasdried over MgSO₄, filtered and evaporated to afford crude product. Thecrude product was purified by flash silica chromatography, elutiongradient 2 to 6% MeOH with ammonia in DCM. Pure fractions wereevaporated to dryness to afford (S)-tert-butyl(4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-yl)methylcarbamate(428 mg, 59.2%) as a colourless gum. The product (428 mg) was dissolvedin dioxane (6.00 mL) and 4M hydrogen chloride in dioxane (0.925 mL,26.64 mmol) added. The reaction was stirred at ambient temperature for 2hours. The crude product was purified by ion exchange chromatography,using an SCX column. The desired product was eluted from the columnusing 3.5N ammonia/MeOH and pure fractions were evaporated to dryness.The crude product was purified by preparative HPLC (Waters XBridge PrepC18 OBD column, 5 μm silica, 19 mm diameter, 100 mm length), usingdecreasingly polar mixtures of water (containing 1% ammonia) and MeCN aseluents. Fractions containing the desired compound were evaporated todryness to afford(S)-4-(aminomethyl)-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(192 mg, 32.5%) as a white dry film.

¹H NMR (400.13 MHz, DMSO-d6) δ 1.45-1.47 (2H, m), 1.79-1.84 (1H, m),1.89-1.99 (1H, m), 2.09 (2H, s), 2.67 (2H, s), 3.36-3.42 (4H, m), 4.25(2H, d), 4.57 (1H, s), 5.01 (1H, d), 6.55 (1H, s), 7.14 (1H, s),7.33-7.36 (4H, m), 8.11 (1H, s), 8.44 (1H, d), 11.61 (1H, s).

MS m/e MH⁺443.

EXAMPLE 24(S)-4-amino-N-(1-(4-bromophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

(S)-3-Amino-3-(4-bromophenyl)propan-1-ol (Intermediate 51) (191 mg, 0.83mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (300 mg, 0.83 mmol) and DIPEA (0.435 mL, 2.49mmol) in DMA (5 mL).O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (347 mg, 0.91 mmol) was added and the resultingsolution was stirred at 20° C. for 24 hours. The reaction mixture wasevaporated to dryness then diluted with EtOAc (300 mL), and washedsequentially with water (50 mL) and saturated brine (50 mL). The organiclayer was dried over MgSO₄, filtered and evaporated to afford crudeproduct. The crude product was purified by flash silica chromatography,elution gradient 2 to 6% MeOH with ammonia in DCM. Fractions wereevaporated to afford (5)-tert-butyl4-(1-(4-bromophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(212 mg, 44.5%) as a white solid. The product (212 mg, 0.36 mmol) wassuspended in dioxane (5.00 mL) and 4M hydrogen chloride in dioxane(0.577 mL, 16.6 mmol) added. The reaction was stirred at ambienttemperature for 2 hours. The crude product was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 3.5N ammonia/MeOH and pure fractions were evaporated todryness. The product was then purified by preparative HPLC (WatersXBridge Prep C18 OBD column, 5 μm silica, 19 mm diameter, 100 mmlength), using decreasingly polar mixtures of water (containing 1%ammonia) and MeCN as eluents. Fractions containing the desired compoundwere evaporated to dryness to afford(S)-4-amino-N-(1-(4-bromophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(51.0 mg, 13.0%) as a white solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 1.38-1.46 (2H, m), 1.79-2.01 (4H, m),2.15 (2H, s), 3.37 (2H, q), 3.51-3.58 (2H, m), 4.37 (2H, t), 4.52 (1H,t), 4.86 (1H, d), 6.57 (1H, d), 7.15 (1H, d), 7.27 (2H, d), 7.48-7.51(2H, m), 8.12 (1H, s), 8.42 (1H, d), 11.62 (1H, s).

MS m/e MH⁺473.

EXAMPLE 254-amino-N-(1-(4-chlorophenyl)-4-(dimethylamino)butyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

1-(4-Chlorophenyl)-N4,N4-dimethylbutane-1,4-diamine (Intermediate 57)(330 mg, 1.46 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (526 mg, 1.46 mmol) and DIPEA (0.763 mL, 4.37mmol) in DMA (5 mL).O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (609 mg, 1.60 mmol) was added and the resultingsolution was stirred at 50° C. for 2 hours. The reaction mixture wasdiluted with EtOAc (25 mL), and washed sequentially with water (20 mL)and saturated brine (20 mL). The organic layer was dried over MgSO₄,filtered and evaporated to afford crude product. The crude product waspurified by flash silica chromatography, elution gradient 5 to 10% MeOHwith ammonia in isohexane. Pure fractions were evaporated to dryness toafford tert-butyl4-(1-(4-chlorophenyl)-4-(dimethylamino)butylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(423 mg, 51.0%) as a colourless gum. tert-Butyl4-(1-(4-chlorophenyl)-4-(dimethylamino)butylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(423 mg, 0.74 mmol) was dissolved in DCM (5.00 mL) and TFA (1 mL) added.The reaction was stirred at ambient temperature for 2 hours. The crudeproduct was purified by ion exchange chromatography, using an SCXcolumn. The desired product was eluted from the column using 3.5Nammonia/MeOH and pure fractions were evaporated to dryness to affordcrude product which was triturated with diethyl ether to afford4-amino-N-(1-(4-chlorophenyl)-4-(dimethylamino)butyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(245 mg, 35.8%) as a white solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 1.26-1.33 (2H, m), 1.38-1.47 (2H, m),1.65-1.75 (2H, m), 1.87-2.01 (2H, m), 2.08 (6H, s), 2.18 (2H, t),3.50-3.58 (2H, m), 4.35-4.41 (2H, m), 4.73 (1H, m), 6.57 (1H, d),7.14-7.16 (1H, m), 7.32-7.37 (4H, m), 8.12 (1H, s), 8.31 (1H, d), 11.62(1H, s).

MS m/e MH⁺470.

EXAMPLE 26(S)-4-amino-N-(1-(4-chlorophenyl)-3-(diethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

(S)-1-(4-Chlorophenyl)-N3,N3-diethylpropane-1,3-diamine (Intermediate60) (119 mg, 0.49 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (179 mg, 0.49 mmol) and DIPEA (0.259 mL, 1.48mmol) in DMA (5 mL).O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (207 mg, 0.54 mmol) was added and the resultingsolution was stirred at 50° C. for 2 hours. The reaction mixture wasdiluted with EtOAc (25 mL), and washed sequentially with water (20 mL)and saturated brine (20 mL). The organic layer was dried over MgSO₄,filtered and evaporated to afford crude product. The crude product waspurified by flash silica chromatography, elution gradient 5 to 10% MeOHwith ammonia in DCM. Pure fractions were evaporated to dryness to afford(S)-tert-butyl4-(1-(4-chlorophenyl)-3-(diethylamino)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(189 mg, 65.5%) as a colourless gum. (S)-tert-butyl4-(1-(4-chlorophenyl)-3-(diethylamino)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(189 mg, 0.33 mmol) was dissolved in DCM (5.00 mL) and TFA (1 mL) added.The reaction was stirred at ambient temperature for 2 hours. The crudeproduct was purified by ion exchange chromatography, using an SCXcolumn. The desired product was eluted from the column using 7Nammonia/MeOH and pure fractions were evaporated to dryness to affordcrude product which was triturated with diethyl ether to afford(S)-4-amino-N-(1-(4-chlorophenyl)-3-(diethylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(70.0 mg, 29.3%) as a white foam.

¹H NMR (400.13 MHz, DMSO-d6) δ 0.94 (6H, d), 1.42-1.49 (2H, m),1.86-2.01 (4H, m), 2.43 (2H, m), 3.51-3.59 (2H, m), 4.37-4.43 (2H, m),4.84 (1H, t), 6.58 (1H, d), 7.15-7.16 (1H, d), 7.32-7.38 (4H, m), 8.12(1H, s), 8.59 (1H, s), 11.63 (1H, s).

MS m/e MH⁺484.

EXAMPLE 27(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(5-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Hydrogen chloride 4M in dioxane (0.923 mL, 3.69 mmol) was added to(5)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(5-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 65) (420 mg, 0.74 mmol) in dioxane (25 mL). The resultingsolution was stirred at ambient temperature for 2 hours. The reactionmixture was dissolved in methanol and purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 3.5N ammonia/MeOH and pure fractions were evaporated todryness. The product was then purified by preparative LCMS (WatersXBridge Prep C18 OBD column, 5μ silica, 19 mm diameter, 100 mm length),using decreasingly polar mixtures of water (containing 1% ammonia) andMeCN as eluents. Fractions containing the desired compound wereevaporated to dryness to afford(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(5-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(25.0 mg, 7.2%) as a white solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 0.66-0.70 (2H, m), 0.86-0.91 (2H, m),1.41-1.50 (2H, m), 1.81-2.13 (5H, m), 3.35-3.43 (4H, m), 3.99-4.07 (2H,m), 4.54 (1H, t), 4.90 (1H, d), 6.90 (1H, s), 7.32-7.38 (4H, m), 8.18(1H, s), 8.47 (1H, d), 11.46 (1H, s).

MS m/e MH⁺470.

EXAMPLE 284-amino-N-(1-(4-chlorophenyl)-3-(methylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Hydrogen chloride (4M in dioxane, 0.461 mL, 1.84 mmol) was added totert-butyl4-(1-(4-chlorophenyl)-3-(methylamino)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 68) (10 mg, 0.02 mmol) in a mixture of DCM (3 mL) andmethanol (1 mL) at 22° C. The resulting solution was stirred at 22° C.for 2 hours. The mixture was evaporated and the residue was purified byion exchange chromatography, using an SCX column. The desired productwas eluted from the column using 30% (2M NH₃ in MeOH) in DCM and purefractions were evaporated to dryness to afford4-amino-N-(1-(4-chlorophenyl)-3-(methylamino)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(7 mg, 86%) as a colourless gum.

¹H NMR (399.902 MHz, DMSO) δ 1.44 (2H, m), 1.83 (2H, dt), 1.87-2.01 (2H,m), 2.15 (2H, m), 2.25 (2H, br.s), 2.42 (2H, m), 3.56 (2H, m), 4.38 (2H,m), 4.84 (1H, br.s), 6.58 (1H, d), 7.16 (1H, d), 7.32-7.38 (4H, m), 8.13(1H, s), 8.57 (1H, s), 11.63 (1H, s).

MS m/e MH⁺442.4.

The compounds of the invention listed in Table A below were made fromthe appropriate starting materials using a process analogous to thatdescribed in Example 2.

TABLE A Compound No. Name Structure NMR/MS 29 4-amino-N-[(4-chlorophenyl) (phenyl)methyl]- 1-(7H- pyrrolo[2,3- d]pyrimidin-4-yl)piperidine-4- carboxamide

1H NMR (399.902 MHz, DMSO) δ 11.65 (1H, s), 8.76 (1H, s), 8.13 (1H, s),7.42- 7.25 (9H, m), 7.17-7.15 (1H, m), 6.60-6.58 (1H, m), 6.07 (1H, s),4.45-4.39 (2H, m), 3.59-3.51 (2H, m), 2.34-2.27 (2H, m), 2.02-1.93 (2H,m), 1.52-1.46 (2H, m) m/z (ESI+) (M + H)+ = 461; HPLC t_(R) = 2.21 min.30 4-amino-N-[2- amino-1-(4- chlorophenyl)-2- oxoethyl]-1-(7H-pyrrolo[2,3- d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (399.902 MHz, DMSO) δ 11.65 (1H, s), 8.92 (1H, s), 8.13 (1H, s),7.80 (1H, s), 7.45-7.39 (4H, m), 7.29 (1H, s), 7.17-7.15 (1H, m),6.58-6.57 (1H, m), 5.30 (1H, s), 4.48-4.38 (2H, m), 3.55- 3.46 (2H, m),2.43 (2H, s), 2.03-1.94 (1H, m), 1.90-1.82 (1H, m), 1.49-1.39 (2H, m)m/z (ESI+) (M + H)+ = 428; HPLC t_(R) = 1.45 min.

The compounds of the invention listed in Table B below were made fromthe appropriate starting materials using a process analogous to thatdescribed in Example 9 alternative route 1.

TABLE B No. Compound Name Structure NMR/MS 31 4-amino-1-(3- bromo-1H-pyrazolo[3,4- d]pyrimidin-4-yl)-N- [(1S)-1-(4- chlorophenyl)ethyl]piperidine-4- carboxamide

MS: m/z (ESI+) (M + H)+ = 480; HPLC tR = 1.34 min. 32 4-amino-N-[(1S)-1-(4- chlorophenyl)ethyl]- 1-(5-chloro-7H- pyrrolo[2,3- d]pyrimidin-4-yl)piperidine-4- carboxamide

¹H NMR (400 MHz, DMSO) δ 1.39 (3H, d), 1.42-1.51 (2H, m), 1.98-2.18 (4H,m), 3.36-3.48 (2H, m), 3.90-4.03 (2H, m), 4.82-4.96 (1H, m), 7.31-7.41(4H, m), 7.45 (1H, s), 8.23 (1H, s), 8.34 (1H, d), 11.98- 12.18 (1H, m).MS: m/z (ESI+) (M + H)+ = 433; HPLC tR = 1.96 min. 33 4-amino-1-(3-bromo-1H- pyrazolo[3,4- d]pyrimidin-4-yl)-N- [(1S)-1-(4-chlorophenyl)-3- hydroxypropyl] piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.45- 1.53 (2H, m), 1.81- 1.94 (2H, m),1.98- 2.14 (2H, m), 3.38 (2H, m), 3.55-3.62 (2H, m), 4.17-4.23 (2H, m),4.54 (1H, t), 4.90 (1H, d), 7.32-7.38 (4H, m), 8.29 (1H, s), 8.46 (1H,d) MS m/e MH+ 510 34 4-amino-N-[(1S)-1- (4-chlorophenyl)-3-hydroxypropyl]-1- (5-chloro-7H- pyrrolo[2,3- d]pyrimidin-4-yl)piperidine-4- carboxamide

1H NMR (399.9 MHz, DMSO-d6) δ 1.42- 1.50 (2H, m), 1.82- 1.92 (2H, m),2.01- 2.15 (2H, m), 3.39- 3.46 (4H, m), 3.93- 3.99 (2H, m), 4.59 (1H,t), 4.90 (1H, m), 7.33- 7.39 (4H, m), 7.47 (1H, s), 8.25 (1H, s), 8.52(1H, d), 12.14 (1H, s) MS m/e MH+ 463 35 4-amino-1-(5- bromo-7H-pyrrolo[2,3- d]pyrimidin-4-yl)-N- [(1S)-1-(4- chlorophenyl)ethyl]piperidine-4- carboxamide

¹H NMR (400 MHz, DMSO) δ 1.38 (3H, d), 1.42-1.52 (2H, m), 2.02-2.23 (4H,m), 3.40-3.45 (2H, m), 3.85-3.99 (2H, m), 4.80-4.97 (1H, m), 7.33-7.40(4H, m), 7.51 (1H, s), 8.26 (1H, s), 8.33 (1H, d), 12.11- 12.28 (1H, m).MS: m/z (ESI+) (M + H)+ = 479; HPLC tR = 2.08 min. 36 4-amino-N-[(1S)-1-(4-chlorophenyl)-3- hydroxypropyl]-1- (5-methyl-7H- pyrrolo[2,3-d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (399.9 MHz, DMSO-d6) δ 1.41- 1.49 (2H, m), 1.79- 2.13 (6H, m),2.34 (3H, s), 3.38-3.42 (2H, m), 3.73 (2H, t), 4.59 (1H, t), 4.90-4.93(1H, m), 7.05 (1H, s), 7.31-7.41 (4H, m), 8.19 (1H, s), 8.50 (1H, d),11.50 (1H, s) MS m/e MH+ 443 37 4-amino-N-[(1S)-1- (4-chlorophenyl)ethyl]- 1-(5-methyl-7H- pyrrolo[2,3- d]pyrimidin-4-yl)piperidine-4- carboxamide

¹H NMR (400 MHz, DMSO) δ 1.39 (3H, d), 1.41-1.54 (2H, m), 1.96-2.17 (4H,m), 2.34 (3H, s), 3.31- 3.43 (2H, m), 3.65- 3.84 (2H, m), 4.81- 4.97(1H, m), 7.03 (1H, s), 7.30-7.44 (4H, m), 8.18 (1H, s), 8.34 (1H, d),11.45 (1H, s). MS: m/z (ESI+) (M + H)+ = 413; HPLC tR = 1.41 min.

The compounds of the invention listed in Table C below were made fromthe appropriate starting materials using a process analogous to thatdescribed in Example 9 alternative route 2.

TABLE C No. Compound Name Structure NMR/MS 38* 4-amino-N-[(1S)-1-(4-chlorophenyl)-3- hydroxy-3- methylbutyl]-1- (7H-pyrrolo[2,3-d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (399.902 MHz, DMSO-d6) δ 1.10 (s, 3H), 1.12 (s, 3H), 1.54- 1.63(m, 2H), 1.66-1.71 (m, 1H), 1.92-1.99 (m, 1H), 2.05-2.16 (m, 2H),3.49-3.60 (m, 2H), 4.41- 4.49 (m, 2H), 4.96 (dd, 1H), 6.61 (d, 1H), 7.18(d, 1H), 7.29-7.35 (m, 4H), 8.15 (s, 1H), 8.62 (d, 1H), 11.64 (s, 1H)m/z (ES+) (M + H)+ = 457.22 39 4-amino-N-[(1S)-1- (4-cyanophenyl)ethyl]- 1-(7H-pyrrolo[2,3- d]pyrimidin-4- yl)piperidine-4-carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.40 (3H, d), 1.51 (2H, d), 1.93- 2.08(2H, m), 3.52-3.58 (2H, m), 4.39-4.45 (2H, m), 4.94 (1H, t), 6.59 (1H,d), 7.16-7.17 (1H, d), 7.51 (2H, d), 7.78 (2H, d), 8.13 (1H, s), 8.47(1H, d), 11.64 (1H, s) MS m/e MH+ 390 40 4-amino-N-[(1S)-1-(3-chlorophenyl)-3- hydroxypropyl]-1- (7H-pyrrolo[2,3- d]pyrimidin-4-yl)piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.44 (2H, t), 1.81-2.03 (4H, m),3.35-3.40 (2H, m), 3.50- 3.58 (2H, m), 4.35- 4.42 (2H, m), 4.54 (1H, t),4.89 (1H, d), 6.58 (1H, m), 7.14-7.16 (1H, m), 7.26-7.28 (2H, m), 7.32-7.36 (2H, m), 8.12 (1H, s), 8.46 (1H, d), 11.62 (1H, s) MS m/e MH+ 42941 4-amino-1-(7H- pyrrolo[2,3- d]pyrimidin-4-yl)- N-{(1S)-1-[4-(trifluoromethyl) phenyl]ethyl} piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.41 (3H, d), 1.50 (2H, d), 1.93- 2.06(2H, m), 3.52-3.58 (2H, m), 4.39-4.44 (2H, m), 4.95 (1H, t), 6.59 (1H,d), 7.15-7.17 (1H, d), 7.54 (2H, d), 7.67 (2H, d), 8.13 (1H, s), 8.46(1H, d), 11.64 (1H, s) MS m/e MH+ 433 *Made by the same HATU couplingmethodology as that described in Example 9 alternative route 2 anddeprotected with TFA using the same methodology as described for Example8.

The compounds of the invention listed in Table D below were made fromthe appropriate starting materials using a process analogous to thatdescribed in Example 16.

TABLE D No. Compound Name Structure NMR/MS 42 4-amino-N-[(1R)-1- (4-bromophenyl)ethyl]- 1-(7H-pyrrolo[2,3- d]pyrimidin-4- yl)piperidine-4-carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.37 (3H, d), 1.39-1.47 (2H, m),1.85-2.02 (2H, m), 2.18 (2H, s), 3.50-3.58 (2H, m), 4.33-4.43 (2H, m),4.80-4.88 (1H, s), 6.57- 6.59 (1H, m), 7.14-7.16 (1H, m), 7.27-7.29 (2H,m), 7.48-7.51 (2H, m), 8.12 (1H, s), 8.30 (1H, d), 11.62 (1H, s) MH+ =443 RT = 1.69 min 43 4-amino-N-[1-(4- chlorophenyl)-2-phenylethyl]-1-(7H- pyrrolo[2,3- d]pyrimidin-4- yl)piperidine-4-carboxamide

1H NMR (400 MHz, DMSO) δ 1.21-1.42 (2H, m), 1.72-1.94 (2H, m), 2.98-3.10(2H, m), 3.44-3.62 (2H, m), 4.13- 4.23 (1H, m), 4.24- 4.35 (1H, m),4.98-5.12 (1H, m), 6.54 (1H, d), 7.12-7.29 (6H, m), 7.32- 7.44 (4H, m),8.12 (1H, s), 8.44 (1H, d), 11.63 (1H, s) MH+ = 475 RT = 2.31 min 444-amino-N-[1-(4- fluorophenyl)ethyl]- 1-(7H-pyrrolo[2,3- d]pyrimidin-4-yl)piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.38 (3H, d), 1.40-1.46 (2H, m),1.87-1.91 (1H, m), 1.96- 2.01 (1H, m), 2.16 (2H, s), 3.50-3.58 (2H, m),4.35-4.43 (2H, m), 4.84- 4.92 (1H, m), 6.57- 6.58 (1H, m), 7.10-7.15(3H, m), 7.34-7.37 (2H, m), 8.12 (1H, s), 8.28 (1H, d), 11.62 (1H, s)MH+ = 383 RT = 1.47 min, 45 4-amino-N-[(4- chlorophenyl)(cyano)methyl]-1-(7H- pyrrolo[2,3- d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (400 MHz, DMSO) δ 1.42-1.58 (3H, m), 1.84-2.04 (2H, m), 3.49-3.64(2H, m), 4.30-4.44 (2H, m), 6.16 (1H, s), 6.57-6.63 (1H, m), 7.13-7.19(1H, m), 7.46-7.55 (4H, m), 8.13 (1H, s), 11.64 (1H, s) MH+ = 410 RT =1.79 min 46 4-amino-N-(1- phenylethyl)-1-(7H- pyrrolo[2,3-d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.38 (3H, d), 1.89-2.03 (2H, m), 2.17(2H, s), 3.50-3.57 (2H, m), 4.37-4.43 (2H, m), 4.84-4.92 (1H, m),6.57-6.58 (1H, m), 7.15 (1H, dd), 7.19-7.26 (1H, m), 7.31-7.32 (4H, m),8.12 (1H, s), 8.28 (1H, d), 11.62 (1H, s) MH+ = 365 RT = 1.40 min,

The compounds of the invention listed in Table E below were made fromthe appropriate starting materials using a process analogous to thatdescribed in Example 25.

TABLE E No. Compound Name Structure NMR/MS 47 4-amino-N-[1-(4-chlorophenyl)-4- pyrrolidin-1- ylbutyl]-1-(7H- pyrrolo[2,3-d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.31-1.49 (4H, m), 1.65 (4H, s),1.69-1.77 (2H, m), 1.88- 1.98 (2H, m), 2.15 (2H, s), 2.34 (4H, s), 2.36(2H, s), 3.53-3.58 (2H, m), 4.34-4.41 (2H, m), 4.73 (1H, m), 6.57 (1H,d), 7.14-7.15 (1H, d), 7.32- 7.37 (4H, m), 8.12 (1H, s), 8.30 (1H, d),11.62 (1H, s) MS m/e MH+ 496 48 4-amino-N-[1-(4- chlorophenyl)-4-morpholin-4- ylbutyl]-1-(7H- pyrrolo[2,3- d]pyrimidin-4-yl)piperidine-4- carboxamide

1H NMR (399.9 MHz, DMSO-d6) δ 1.32-1.37 (1H, m), 1.44-1.51 (1H, m),1.57-1.76 (4H, m), 2.10-2.17 (2H, m), 2.26- 2.35 (6H, m), 3.47- 3.56(6H, m), 4.51 (2H, d), 4.79 (1H, m), 6.62- 6.64 (1H, m), 7.19 (1H, t),7.32-7.34 (2H, d), 7.37-7.39 (2H, d), 8.16 (1H, s), 8.49 (1H, s), 11.68(1H, s) MS m/e MH+ 512 49 4-amino-N-[1-(4- chlorophenyl)-4-piperidin-1-ylbutyl]- 1-(7H-pyrrolo[2,3- d]pyrimidin-4- yl)piperidine-4-carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.31-1.37 (3H, m), 1.41-1.47 (6H, m),1.66-1.73 (2H, m), 1.86-2.00 (2H, m), 2.16- 2.24 (7H, m), 3.50- 3.58(2H, m), 4.34-4.40 (2H, m), 4.73 (1H, m), 6.57 (1H, d), 7.15 (1H, d),7.31-7.37 (4H, m), 8.12 (1H, s), 8.29 (1H, d), 11.62 (1H, s) MS m/e MH+510

EXAMPLE 49A AND 49B4-amino-N-[(1S)-1-(4-chlorophenyl)-4-piperidin-1-ylbutyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideand4-amino-N-[(1R)-1-(4-chlorophenyl)-4-piperidin-1-ylbutyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

The individual stereoisomers of the racemate of Example 49 wereseparated using the same methodology as that described for Examples 11Aand 11B.

1H NMR (500.13 MHz, DMSO-d6) δ 1.31-1.41 (4H, m), 1.44-1.48 (6H, m),1.68-1.71 (2H, m), 1.90-1.98 (4H, m), 2.21 (6H, m), 3.54-3.56 (2H, m),4.38 (2H, t), 4.73 (1H, d), 6.58 (1H, q), 7.15 (1H, q), 7.32-7.37 (4H,m), 8.12 (1H, s), 8.29 (1H, s), 11.63 (1H, s)

MS m/e MH+510

1H NMR (500.13 MHz, DMSO-d6) δ 1.31-1.41 (4H, m), 1.44-1.48 (7H, m),1.68-1.71 (2H, m), 1.90-1.98 (2H, m), 2.21 (6H, m), 3.54-3.56 (2H, m),4.38 (2H, t), 4.73 (1H, d), 6.58 (1H, q), 7.15 (1H, q), 7.32-7.37 (4H,m), 8.12 (1H, s), 8.29 (1H, s), 11.63 (1H, s)

MS m/e MH+510

The compounds of the invention listed in Table F below were made fromthe appropriate starting materials using a process analogous to thatdescribed in Example 26.

TABLE F No. Compound Name Structure NMR/MS 50 4-amino-N-[(1S)-1-(4-chlorophenyl)-3- (4-methylpiperazin- 1-yl)propyl]-1-(7H- pyrrolo[2,3-d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.39-1.47 (2H, m), 1.84-1.97 (4H, m),2.13 (4H, s), 2.19 (2H, t), 2.31-2.34 (7H, m), 3.54-3.57 (2H, m), 4.38(2H, m), 4.82 (1H, m), 6.57 (1H, d), 7.14- 7.16 (1H, d), 7.31-7.37 (4H,m), 8.12 (1H, s), 8.55 (1H, d), 11.62 (1H, s) MS m/e MH+ 511 514-amino-N-[(1S)-1- (4-chlorophenyl)-3- morpholin-4- ylpropyl]-1-(7H-pyrrolo[2,3- d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.43 (2H, m), 1.83-2.00 (4H, m), 2.20(2H, t), 2.27-2.37 (4H, m), 3.51-3.63 (6H, m), 4.35-4.41 (2H, m), 4.85(1H, m), 6.57 (1H, d), 7.15 (1H, d), 7.32- 7.37 (4H, m), 8.12 (1H, s),8.65 (1H, d), 11.62 (1H, s) MS m/e MH+ 498 52 4-amino-N-[(1S)-1-(4-chlorophenyl)-3- piperidin-1- ylpropyl]-1-(7H- pyrrolo[2,3-d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.33-1.50 (8H, m), 1.82-1.90 (4H, m),2.15 (2H, t), 2.25- 2.34 (4H, m), 3.53-3.57 (2H, m), 4.39 (2H, m), 4.82(1H, m), 6.57 (1H, d), 7.14-7.16 (1H, d), 7.30-7.37 (4H, m), 8.12 (1H,s), 8.64-8.66 (1H, d), 11.62 (1H, s) MS m/e MH+ 496 53*4-amino-N-[(1S)-1- (4-chlorophenyl)-3- piperazin-1- ylpropyl]-1-(7H-pyrrolo[2,3- d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.40-1.47 (2H, m), 1.82-1.99 (4H, m),2.11-2.25 (6H, m), 2.67-2.69 (4H, m), 3.54- 3.57 (2H, m), 4.35- 4.41(2H, m), 4.83 (1H, m), 6.57 (1H, d), 7.15 (1H, d), 7.31-7.37 (4H, m),8.12 (1H, s), 8.61 (1H, d), 11.62 (1H, s) MS m/e MH+ 497 544-amino-N-[(1S)-1- (4-chlorophenyl)-3- (1H-imidazol-1- yl)propyl]-1-(7H-pyrrolo[2,3- d]pyrimidin-4- yl)piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.41-1.53 (2H, m), 1.86-2.04 (2H, m),2.13-2.25 (4H, m), 3.52-3.61 (2H, m), 3.96- 4.00 (2H, m), 4.34- 4.42(2H, m), 4.69 (1H, m), 6.58 (1H, d), 6.89 (1H, s), 7.15 (2H, t), 7.31(2H, d), 7.36-7.38 (2H, d), 7.56 (1H, s), 8.12 (1H, s), 8.44 (1H, d),11.62 (1H, s) MS m/e MH+ 479 55 4-amino-N-[(1S)-1- (4-chlorophenyl)-3-pyrrolidin-1- ylpropyl]-1-(7H- pyrrolo[2,3- d]pyrimidin-4-yl)piperidine-4- carboxamide

1H NMR (400.13 MHz, DMSO-d6) δ 1.42-1.58 (2H, m), 1.71 (5H, s),1.84-2.02 (5H, m), 2.33 (2H, m), 2.55 (2H, m), 3.51-3.59 (2H, m), 4.36-4.43 (2H, m), 4.86 (1H, t), 6.58 (1H, d), 7.16 (1H, d), 7.30-7.38 (5H,m), 8.12 (1H, s), 11.63 (1H, s) MS m/e MH+ 482 *Made by the samemethodology as that described for Example 26 starting from1-(2,4-dimethoxybenzyl)piperazine. The TFA deprotection in the finalstep was heated at 80° C. to remove Boc and dimethoxybenzyl groups.

EXAMPLE 564-amino-N-(1-(4-chlorophenyl)-2-sulfamoylethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Tert-butyl4-(1-(4-chlorophenyl)-2-(methylsulfonamido)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 76) (100 mg, 0.17 mmol) was treated with trifluoroaceticacid (2 mL). The solution was stirred for 1 hour at room temperature.The mixture was concentrated under reduced pressure. The crude productwas purified by ion exchange chromatography, using an SCX column. Theresidue was loaded onto the column in methanol and washed with methanol.The desired product was eluted from the column using 2M ammonia inmethanol and pure fractions were evaporated to dryness to afford4-amino-N-(1-(4-chlorophenyl)-2-(methylsulfonamido)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(79 mg, 95%) as a colourless solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.38-1.53 (2H, m), 1.85-2.07 (2H, m), 2.20(2H, br, s), 2.85 (3H, s), 3.57 (2H, m), 4.34-4.46 (2H, m), 4.87-4.94(1H, m), 6.57-6.60 (1H, m), 7.12-7.19 (2H, m), 7.35-7.43 (4H, m), 8.13(1H, s), 8.46 (1H, br, s), 11.64 (1H, s)

MS m/e MH⁺=492

EXAMPLE 574-amino-N-(1-(4-chlorophenyl)-2-sulfamoylethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Tert-butyl4-(1-(4-chlorophenyl)-2-sulfamoylethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 81) (153 mg, 0.26 mmol) was treated with trifluoroaceticacid (4 mL). The solution was stirred for 30 minutes at roomtemperature. The mixture was concentrated under reduced pressure and theresidue was purified by ion exchange chromatography, using an SCXcolumn. The desired product was eluted from the column using ammonia inmethanol (2M) and pure fractions were evaporated to dryness to afford4-amino-N-(1-(4-chlorophenyl)-2-sulfamoylethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(125 mg, 99%) as a colourless solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.35-1.53 (2H, m), 1.86-2.04 (2H, m),3.35-3.40 (1H, m), 3.52-3.62 (2H, m), 3.68 (1H, dd), 4.33-4.41 (2H, m),5.24-5.29 (1H, m), 6.56-6.60 (1H, m), 6.88 (2H, s), 7.13-7.17 (1H, m),7.39 (4H, s), 8.13 (1H, s), 8.68 (1H, br, s), 11.63 (1H, s)

MS m/e MH⁺=478

EXAMPLE 58N-(2-acetamido-1-(4-chlorophenyl)ethyl)-4-amino-1-(7H-pyrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Tert-butyl4-(2-acetamido-1-(4-chlorophenyl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 84) (97 mg, 0.17 mmol) was treated with trifluoroaceticacid (2 mL). The solution was stirred for 1 hours at room temperature.The mixture was concentrated under reduced pressure. The crude productwas purified by ion exchange chromatography, using an SCX column. Theresidue was loaded onto the column in methanol and washed with methanol.The desired product was eluted from the column using 2M ammonia inmethanol and pure fractions were evaporated to dryness to affordN-(2-acetamido-1-(4-chlorophenyl)ethyl)-4-amino-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(80 mg, quant.) as a cream dry film.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.43 (2H, t), 1.79 (3H, s), 1.83-2.04 (2H,m), 2.20 (2H, br, s), 3.32-3.38 (2H, m), 3.58 (2H, q), 4.32-4.42 (2H,m), 4.82-4.88 (1H, m), 6.56-6.60 (1H, m), 7.14-7.18 (1H, m), 7.33 (2H,d), 7.38 (2H, d), 7.94 (1H, t), 8.13 (1H, s), 8.42-8.50 (1H, m), 11.63(1H, s)

MS m/e MH⁺=456

EXAMPLE 594-amino-N-(1-(4-chlorophenyl)-2-(1H-imidazol-2-yl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Trifluoroacetic acid (5 mL, 64.90 mmol) was added to tert-butyl4-(1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 89) (310 mg, 0.38 mmol) at room temperature and theresulting solution stirred for 30 minutes. Water (5-6 drops) was thenadded and stirring continued for a further 30 minutes. The resultingsolution was diluted with methanol and applied to a 10 g SCX columnwhich was then eluted with MeOH followed by 2N NH₃ (in MeOH).Product-containing fractions were combined and concentrated byevaporation then triturated with MeCN/DMF (1:1) to give a pale yellowppt. The precipitate was collected by filtration, washed with MeCN anddried under vacuum to afford4-amino-N-(1-(4-chlorophenyl)-2-(1H-imidazol-2-yl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(130 mg, 72.8%) as a cream solid.

1H NMR (399.902 MHz, DMSO) δ 1.36-1.40 (2H, m), 1.85-1.93 (2H, m), 3.08(2H, d), 3.55 (2H, m), 4.29-4.35 (2H, m), 5.18 (1H, q), 6.57 (1H, d),6.85 (2H, s), 7.16 (1H, d), 7.25 (2H, d), 7.31 (2H, d), 8.12 (1H, s),8.89 (1H, d), 11.63 (1H, br s) m/z (ESI+) (M+H)+=465; HPLC tR=1.62 min.

EXAMPLE 604-amino-N-[1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

TFA (0.7 mL) was added to a suspension of tert-butyl4-(1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(180 mg, 0.32 mmol) in dichloromethane (Intermediate 93) (7 mL) underargon. The resulting solution was stirred at room temperature overnight.The solvents were removed in vacuo and the reaction mixture was purifiedby preparative HPLC using a Waters X-Bridge reverse-phase column (C-18,5 microns silica, 19 mm diameter, 100 mm length) and decreasingly polarmixtures of water (containing 0.2% ammonium carbonate) and acetonitrileas eluent. The fractions were evaporated to dryness to afford4-amino-N-[1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamideas a white powder.

1H NMR (500 MHz, DMSO-d6) δ 1.33 (2H, m), 1.83 (2H, m), 2.14 (2H, s),3.53 (2H, m), 4.28 (2H, m), 4.43 (2H, m), 5.22 (1H, d), 6.15 (1H, t),6.56 (1H, q), 7.15 (1H, t), 7.30 (2H, d), 7.36 (2H d), 7.44 (1H, d),7.55 (1H, d), 8.11 (1H, s), 8.81 (1H, br d), 11.64 (1H, s);

m/z (ESI+) (M+H)+=465.

EXAMPLE 614-amino-N-[1-(4-chlorophenyl)-2-(3-methylisoxazol-5-yl)ethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

4-Amino-N-[1-(4-chlorophenyl)-2-(3-methylisoxazol-5-yl)ethyl]-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamidewas made from the appropriate starting materials using a processanalogous to that described for Example 60.

1H NMR (500 MHz, DMSO-d6) δ 1.35 (2H, d), 1.87 (2H, m), 2.14 (2H, s),2.15 (3H, s), 3.17-3.33 (2H, m), 3.53 (2H, m), 4.31 (2H, m), 5.15 (1H,d), 6.06 (1H, s), 6.56 (1H, d), 7.16 (1H, t), 7.39 (4H, m), 8.12 (1H,s), 8.60 (1H, br d), 11.65 (1H, s). m/z (ESI+) (M+H)+=480

EXAMPLE 624-amino-N-(1-(4-chlorophenyl)-2-(thiazol-2-yl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

TFA (2.0 mL) was added to a suspension of tert-butyl4-(1-(4-chlorophenyl)-2-(thiazol-2-yl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 96) (918 mg, 0.95 mmol) in dichloromethane (20 mL) underargon. The resulting solution was stirred at room temperature for onenight. The solvents were removed in vacuo and the reaction mixture waspurified by preparative HPLC using a Waters X-Bridge reverse-phasecolumn (C-18, 5 microns silica, 19 mm diameter, 100 mm length) anddecreasingly polar mixtures of water (containing 0.2% ammoniumcarbonate) and acetonitrile as eluent. The fractions were evaporated todryness to afford4-amino-N-(1-(4-chlorophenyl)-2-(thiazol-2-yl)ethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(259 mg, 56.8%) as a white powder.

1H NMR (500 MHz, DMSO-d6) δ 1.35 (2H, d), 1.85 (2H, m), 2.07 (2H, br s),3.41-3.56 (4H, m), 4.30 (2H, m), 5.20 (1H, d), 6.56 (1H, d), 7.15 (1H,t), 7.36 (4H, m), 7.54 (1H, d), 7.70 (1H, d), 8.11 (1H, s), 8.75 (1H, brd), 11.70 (1H, br s);

m/z (ESI+) (M+H)+=482

EXAMPLE 634-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)-3-oxopropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

TFA (1.0 mL) was added to a suspension of tert-butyl4-(1-(4-chlorophenyl)-3-(dimethylamino)-3-oxopropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 98) (770 mg, 0.24 mmol) in dichloromethane (10 mL) underargon. The resulting solution was stirred at room temperature for onenight. The solvents were removed in vacuo and the reaction mixture waspurified by preparative HPLC using a Waters X-Bridge reverse-phasecolumn (C-18, 5 microns silica, 19 mm diameter, 100 mm length) anddecreasingly polar mixtures of water (containing 0.2% ammoniumcarbonate) and acetonitrile as eluent. The fractions were evaporated todryness to afford4-amino-N-(1-(4-chlorophenyl)-3-(dimethylamino)-3-oxopropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(74.0 mg, 64.8%) as a white powder.

1H NMR (500 MHz, DMSO-d6) δ 1.43 (2H, m), 1.94 (2H, m), 2.75 (3H, s),2.8 (2H, m), 2.89 (3H, s), 3.52 (2H, m), 4.40 (2H, m), 5.15 (1H, d),6.58 (1H, d), 7.16 (1H, t), 7.35 (4H, s), 8.12 (1H, s), 8.76 (1H, br d),11.66 (1H, br s);

m/z (ESI+) (M+H)+=470

EXAMPLE 644-amino-N-(1-(4-chlorophenyl)-3-methoxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Hydrogen chloride (4M in 1,4-dioxane, 1.151 mL, 4.60 mmol) was added totert-butyl4-(1-(4-chlorophenyl)-3-methoxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 101) (50 mg, 0.09 mmol) in a mixture of DCM (5 mL) andmethanol (2 mL) at 22° C. The resulting solution was stirred at 22° C.for 2 days. The mixture was evaporated and the residue was purified byion exchange chromatography, using an SCX column. The desired productwas eluted from the column using 2M NH₃/MeOH; pure fractions wereevaporated to dryness and the residue was triturated with diethyl etherto afford4-amino-N-(1-(4-chlorophenyl)-3-methoxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(39.0 mg, 96%) as a white solid.

1H NMR (399.902 MHz, DMSO) δ 1.44 (2H, m), 1.88-2.02 (5H, m), 2.46 (2H,s), 3.21 (3H, s), 3.28 (2H, t), 3.55 (2H, m), 4.39 (2H, m), 4.87 (1H,dt), 6.59 (1H, dd), 7.16 (1H, dd), 7.33 (2H, d), 7.37 (2H, d), 8.13 (1H,s), 8.45 (1H, d), 11.63 (1H, s);

m/z (ESI+) (M+H)+=443.4; HPLC tR=1.87 min.

EXAMPLE 654-amino-N-(1-(4-chlorophenyl)-3-sulfamoylpropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

Hydrogen chloride (4M in dioxane, 0.676 mL, 2.70 mmol) was added totert-butyl4-(1-(4-chlorophenyl)-3-sulfamoylpropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 105) (16 mg, 0.03 mmol) in a mixture of DCM (5 mL) andmethanol (5 mL) at 22° C. The resulting solution was stirred at 22° C.for 20 hours. The mixture was evaporated and the residue was purified byion exchange chromatography, using an SCX column. The desired productwas eluted from the column using 30% (2M NH₃ in MeOH) in DCM and purefractions were evaporated to dryness to afford4-amino-N-(1-(4-chlorophenyl)-3-sulfamoylpropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(12.00 mg, 90%) as a colourless gum.

1H NMR (399.902 MHz, DMSO) δ 1.48 (2H, m), 1.90-2.06 (2H, m), 2.09-2.24(2H, m), 2.87 (1H, ddd), 3.02 (1H, ddd), 3.56 (2H, m), 3.56 (2H, d),4.41 (2H, m), 4.91 (1H, br.s), 6.59 (1H, dd), 6.80 (2H, s), 7.16 (1H,dd), 7.38-7.43 (4H, m), 8.13 (1H, s), 8.46 (1H, s), 11.64 (1H, s);

m/z (ESI+) (M+H)+=492.4, 494.3; HPLC tR=1.60 min.

EXAMPLE 664-amino-N-(3-amino-1-(4-chlorophenyl)-3-oxopropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

HCl in 1,4-dioxane (4M) (0.228 mL, 0.91 mmol) was added to tert-butyl4-(3-amino-1-(4-chlorophenyl)-3-oxopropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 108) (99 mg, 0.18 mmol) in DCM (4 mL) at 20° C. Theresulting solution was stirred at 20° C. for 3 hours. The reactionmixture was filtered through a PTFE cup and the collected solid waspurified by preparative HPLC (Waters XTerra C18 column, 5μ silica, 30 mmdiameter, 100 mm length), using decreasingly polar mixtures of water(containing 1% NH₃) and MeCN as eluents. Fractions containing thedesired compound were evaporated to dryness to afford4-amino-N-(3-amino-1-(4-chlorophenyl)-3-oxopropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(9.00 mg, 11.1%) as a cream solid.

1H NMR (399.9 MHz, DMSO-d6) δ 1.40-1.43 (2H, m), 1.93-1.96 (2H, m), 2.18(2H, s), 3.54-3.56 (2H, m), 4.36-4.40 (2H, m), 5.12 (1H, d), 6.58 (1H,d), 6.81 (1H, s), 7.15-7.16 (1H, m), 7.32-7.37 (5H, m), 8.13 (1H, s),8.76 (1H, d), 11.63 (1H, s), (no NH₂ visible).

m/z (ESI+) (M+H)+=442; HPLC tR=1.47 min.

EXAMPLE 674-amino-N-(1-(4-chlorophenyl)-3-ureidopropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

TFA (3 mL) was added to tert-butyl4-(1-(4-chlorophenyl)-3-ureidopropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 112) (379 mg, 0.66 mmol). The reaction was left to stir at20° C. for 4 hours and then vacuumed to dryness. The crude product waspurified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using 0.35M NH₃/MeOH and purefractions were evaporated to dryness to afford crude material. The crudeproduct was purified by preparative HPLC (Waters XBridge Prep C18 OBDcolumn, 5μ silica, 19 mm diameter, 100 mm length), using decreasinglypolar mixtures of water (containing 1% NH₃) and MeCN as eluents.Fractions containing the desired compound were evaporated to dryness toafford4-amino-N-(1-(4-chlorophenyl)-3-ureidopropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(14.00 mg, 5.60%) as a white solid.

1H NMR (399.9 MHz, DMSO-d6) δ 1.41-1.49 (2H, m), 1.78-1.88 (2H, m), 2.01(1H, d), 2.91 (1H, t), 2.97 (2H, t), 3.55 (2H, d), 4.37 (2H, d), 4.78(1H, d), 5.40 (2H, s), 5.95 (1H, t), 6.58-6.59 (1H, m), 7.15-7.16 (1H,m), 7.33-7.39 (4H, m), 8.13 (1H, s), 8.36 (1H, d), 11.63 (1H, s);

m/z (ESI+) (M+H)+=471; HPLC tR=1.50 min.

EXAMPLE 684-amino-N-(1-(4-chlorophenyl)-2-cyanoethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

HCl (4M) in 1,4-dioxane (2.011 mL, 8.04 mmol) was added to tert-butyl4-(1-(4-chlorophenyl)-2-cyanoethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 114) (0.281 g, 0.54 mmol) in DCM (8 mL) at 20° C. Theresulting solution was stirred at 20° C. for 18 hours. The reaction wasvacuumed to dryness and was purified by preparative HPLC (Waters XTerraC18 column, 5μ silica, 19 mm diameter, 100 mm length), usingdecreasingly polar mixtures of water (containing 1% NH₃) and MeCN aseluents. Fractions containing the desired compound were evaporated todryness to afford4-amino-N-(1-(4-chlorophenyl)-2-cyanoethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(0.096 g, 42.2%) as a white solid.

1H NMR (399.9 MHz, DMSO-d6) δ 1.42-1.46 (2H, m), 1.52 (2H, d), 1.95 (1H,d), 1.98-2.01 (1H, m), 3.08 (1H, t), 3.56-3.59 (2H, m), 4.38-4.42 (2H,m), 5.18 (1H, s), 6.59-6.60 (1H, m), 7.16 (1H, t), 7.44 (4H, m), 8.13(1H, s), 11.65 (1H, s), (no Visible NH2),

m/z (ESI+) (M+H)+=424; HPLC tR=1.82 min.

EXAMPLE 694-amino-N-(1-(4-chlorophenyl)-3-(methylsulfonamido)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

HCl (4M) in 1,4-Dioxane (1.615 mL, 6.46 mmol) was added to tert-butyl4-(1-(4-chlorophenyl)-3-(methylsulfonamido)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 117) (261 mg, 0.43 mmol), The resulting suspension wasstirred at 20° C. for 1 hour. No reaction. TFA (1 mL) was then added tothe reaction and stirred for 16 hours. The reaction was vacuumed todryness and was purified by ion exchange chromatography, using an SCXcolumn. The desired product was eluted from the column using 0.35MNH₃/MeOH and pure fractions were evaporated to dryness to afford crudematerial. The crude product was purified by preparative HPLC (WatersXBridge Prep C18 OBD column, 5μ silica, 19 mm diameter, 100 mm length),using decreasingly polar mixtures of water (containing [AP-HPLC Buffer])and MeCN as eluents. Fractions containing the desired compound wereevaporated to dryness to afford4-amino-N-(1-(4-chlorophenyl)-3-(methylsulfonamido)propyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(142 mg, 65.2%) as a white solid.

1H NMR (399.9 MHz, DMSO-d6) δ 1.40-1.48 (2H, m), 1.86-1.90 (2H, m),1.93-1.97 (2H, m), 2.17 (2H, s), 2.88 (3H, s), 2.93-2.97 (2H, m),3.53-3.60 (2H, m), 4.37 (2H, t), 4.87 (1H, d), 6.57-6.59 (1H, m), 7.00(1H, t), 7.15-7.16 (1H, m), 7.35-7.40 (4H, m), 8.13 (1H, s), 8.38 (1H,d), 11.63 (1H, s),

m/z (ESI+) (M+H)+=506; HPLC tR=1.62 min.

Intermediate 1:4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid

To a mixture of4-[(2-methylpropan-2-yl)oxycarbonylamino]piperidine-4-carboxylic acid(115.6 g) in CH₃CN—H₂O (6 L) was added to NaHCO₃ (181 g), followed by4-chloro 7H-pyrrole[2,3-d]pyrimidine (72.7 g). The mixture was heated toreflux overnight under nitrogen for 24 hrs and then extracted with EtOAc(1 L×4). The aqueous layer was concentrated and MeOH (1.5 L) was added.The mixture was shaken for 30 min at 45° C. and filtered. The filtratewas concentrated again and dissolved in H₂O (300 mL). 6N HCl was addeduntil the pH reached 4.5 (ca. 80 mL). The mixture was filtered and thecake was dried under vacuum to afford the crude product, which wasfurther purified by silica gel chromatography (eluting withMeOH:DCM=1:3) to yield4-[(2-methylpropan-2-yl)oxycarbonylamino]-1-(3,5,7-triazabicyclo[4.3.0]nona-2,4,8,10-tetraen-2-yl)piperidine-4-carboxylicacid as pale grey solid (105 g, 63%).

1H NMR (400.13 MHz, DMSO-d6) δ 1.40 (9H, s), 1.88-1.95 (2H, m),2.02-2.06 (2H, m), 3.44-3.51 (2H, m), 4.30 (2H, d), 6.60-6.61 (1H, m),7.16-7.18 (1H, m), 7.29 (1H, s), 8.14 (1H, s), 11.68 (1H, s).

MS m/e MH⁺362.

Intermediate 2: (4-chlorophenyl)(cyclopropyl)methanone O-methyl oxime

Methoxylamine hydrochloride (2.004 g) was added to(4-chlorophenyl)(cyclopropyl)methanone (2.71 g) in pyridine (60 mL) at25° C. The resulting solution was stirred at 25° C. for 24 hours. Thepyridine was removed in vacuo, and the residual solid extracted withether. Filtration and evaporation of the solvent gave the crude(4-chlorophenyl)(cyclopropyl)methanone O-methyl oxime (2.75 g, 87%) as ayellow oil. This material was used directly in the next step withoutfurther purification.

MS m/e MH⁺210.

Intermediate 3: (4-chlorophenyl)(cyclopropyl)methanamine

Borane tetrahydrofuran complex (1N in THF, 65.6 mL) was added to(4-chlorophenyl)(cyclopropyl)methanone O-methyl oxime (Intermediate 2)(2.75 g) in THF (100 mL) at 25° C. under nitrogen. The resultingsolution was stirred at reflux for 3 hours, then cooled to 0° C. Waterwas carefully added, followed by aqueous NaOH (20%, 100 mL). Theresulting mixture was stirred at reflux overnight, then allowed to coolto room temperature. The product was extracted into hexane, then driedover sodium sulfate, filtered and evaporated to afford(4-chlorophenyl)(cyclopropyl)methanamine as a clear oil (2.205 g, 93%).

¹H NMR (400.13 MHz, CDCl₃) δ 0.24-0.29 (1H, m), 0.30-0.13 (1H, m),0.45-0.50 (1H, m), 0.58-0.64 (1H, m), 1.01-1.09 (1H, m), 1.77 (2H, s),3.20 (1H, d), 7.28-7.32 (2H, m), 7.33-7.36 (2H, m).

Intermediate 4: 2-amino-2-(4-chlorophenyl)acetonitrile

Lithium bis(trimethylsilyl)amide (42.7 mL) was added to4-chlorobenzaldehyde (5 g) in THF (100 mL) at −40° C. under nitrogen.The resulting solution was warmed to room temperature and stirred for 4hours. α-Hydroxyisobutyronitrile (acetone cyanohydrin, 6.50 mL) was thenadded, and the reaction mixture was stirred at 25° C. for a further 12hours, then quenched with saturated NaHCO₃ (50 mL), extracted with EtOAc(3×100 mL), the organic layer was dried over MgSO₄, filtered andevaporated. The crude material was then purified by flash silicachromatography, elution gradient 0 to 100% EtOAc in isohexane. Purefractions were evaporated to dryness to afford2-amino-2-(4-chlorophenyl)acetonitrile as a colourless oil, whichsolidified on standing to give a white solid (3.40 g, 57.4%). 1H NMR(400.13 MHz, CDCl₃) δ 1.85 (2H, s), 4.82 (1H, s), 7.30-7.34 (2H, m),7.39-7.43 (2H, m).

Intermediate 5: tert-butyl44(4-chlorophenyl)(cyano)methylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU, 0.456 g) was added to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (0.361 g), 2-amino-2-(4-chlorophenyl)acetonitrile(Intermediate 4) (0.167 g) and N-ethyldiisopropylamine (0.523 mL, 3.00mmol) in DMA (5 mL) at 25° C. The resulting solution was stirred at 50°C. for 1 hour. The crude product was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 7M NH₃/MeOH and pure fractions were evaporated todryness to afford tert-butyl4-((4-chlorophenyl)(cyano)methylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamateas a white solid (0.343 g, 67.3%).

MS m/e MH⁺510.

Intermediate 6: tert-butyl4-(2-amino-1-(4-chlorophenyl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

Raney™ nickel (0.257 g), was added to tert-butyl4-((4-chlorophenyl)(cyano)methylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 5) (0.510 g) in ethanol (30 mL). Ammonium hydroxide (0.039mL) was added. This mixture was placed under a balloon of hydrogen andstirred for 48 hours. The reaction mixture was filtered through celiteand the solvent evaporated to dryness. The reaction mixture wasevaporated to dryness and redissolved in DCM (200 mL), and washed withwater (125 mL). The organic layer was dried over MgSO₄, filtered andevaporated to afford crude product, tert-butyl4-(2-amino-1-(4-chlorophenyl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamateas a colourless gum (0.514 g, 100%). This material was used crude in thenext step without further purification.

MS m/e MH⁺514.

Intermediate 7: 1-tert-butyl 4-ethyl 4-cyanopiperidine-1,4-dicarboxylate

A solution of LDA (107 ml, 214.01 mmol) was added to a stirred solutionof tert-butyl 4-cyanopiperidine-1-carboxylate (30 g, 143 mmol) in THF(250 ml) at −78° C., under nitrogen. The resulting solution was stirredat −78° C. for 30 minutes. Ethyl chloroformate (16.37 ml, 171.2 mmol)was added. The resulting solution was stirred and allowed to warm toroom temperature. The reaction mixture was quenched with saturatedNaHCO₃ (250 ml), extracted with DCM, and the organic layer was washedwith saturated brine (100 ml) then dried over MgSO₄, filtered andevaporated to afford the crude material as a orange oil. This materialwas purified by flash silica chromatography, elution gradient 10% EtOAcin isohexane. Pure fractions were evaporated to dryness to afford1-tert-butyl 4-ethyl 4-cyanopiperidine-1,4-dicarboxylate (20.8 g, 51.6%)as a yellow oil.

1H NMR (400.13 MHz, CDCl₃) δ 1.33 (3H, t), 1.46 (9H, s), 1.96-2.00 (2H,m), 2.04-2.08 (2H, m), 3.12 (2H, s), 4.09-4.14 (2H, m), 4.29 (2H, q).

Intermediate 8: 1-tert-butyl 4-ethyl4-(aminomethyl)piperidine-1,4-dicarboxylate

Platinum (IV) oxide (0.724 g, 3.19 mmol) and 1-tert-butyl 4-ethyl4-cyanopiperidine-1,4-dicarboxylate (Intermediate 7) (9 g, 31.9 mmol) inacetic acid (100 ml) were stirred under an atmosphere of hydrogen at 5bar and 25° C. for 1 day. The crude product was filtered through celiteand the filtrate purified by ion exchange chromatography, using an SCXcolumn. The desired product was eluted from the column using 7M NH₃/MeOHand pure fractions were evaporated to dryness to afford 1-tert-butyl4-ethyl 4-(aminomethyl)piperidine-1,4-dicarboxylate (7.59 g, 83%) as acolourless oil.

1H NMR (400.13 MHz, CDCl₃) δ 1.27-1.28 (3H, m), 1.30-1.37 (2H, m), 1.41(2H, s), 1.45 (9H, s), 2.10 (2H, d), 2.78 (2H, s), 2.91-2.97 (2H, m),3.89 (2H, s), 4.21 (2H, q).

Intermediate 9: ethyl 4-(aminomethyl)piperidine-4-carboxylate

Hydrogen chloride 4M in dioxane (33.2 ml, 132.7 mmol) was added to1-tert-butyl 4-ethyl 4-(aminomethyl)piperidine-1,4-dicarboxylate(Intermediate 8) (7.6 g, 26.5 mmol) in dioxane (35 ml). The resultingsolution was stirred at 20° C. for 3 hours. The crude product waspurified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using 7M NH₃/MeOH and purefractions were evaporated to dryness to afford ethyl4-(aminomethyl)piperidine-4-carboxylate (3.34 g, 67.6%) as a yellowliquid.

1H NMR (400.13 MHz, CDCl₃) δ 1.23-1.30 (3H, m), 1.26-1.37 (2H, m), 2.12(2H, d), 2.65-2.72 (2H, m), 2.77 (2H, s), 2.94-2.99 (2H, m), 4.21 (2H,q).

Intermediate 10: ethyl4-(aminomethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylate

N-Ethyldiisopropylamine (3.70 ml, 21.26 mmol) was added to ethyl4-(aminomethyl)piperidine-4-carboxylate (Intermediate 9) (3.3 g, 17.7mmol) and 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (2.72 g, 17.72 mmol) inDMA (35 ml). The resulting solution was stirred at 60° C. for 18 hours.The crude product was purified by ion exchange chromatography, using anSCX column. The desired product was eluted from the column using 7MNH₃/MeOH and pure fractions were evaporated to dryness to afford ethyl4-(aminomethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylate(5.08 g, 95%) as a beige solid.

1H NMR (400.13 MHz, DMSO-d6) δ 1.22 (3H, t), 1.44-1.51 (2H, m),2.04-2.07 (2H, m), 2.67 (2H, d), 3.23-3.30 (2H, m), 4.15 (2H, q),4.39-4.44 (2H, m), 6.59 (1H, t), 7.16-7.17 (1H, m), 8.12 (1H, s), 11.67(1H, s)

MS m/e MH⁺304.

Intermediate 11: ethyl4-((tert-butoxycarbonylamino)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylate

Di-tert-butyl dicarbonate (470 mg, 2.15 mmol) was added to ethyl4-(aminomethyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylate(Intermediate 10) (653 mg, 2.15 mmol) and triethylamine (0.300 ml, 2.15mmol) in DCM (10 ml). The resulting suspension was stirred at ambienttemperature for 2 hours. The reaction mixture was diluted with DCM (50mL), and washed sequentially with water (50 mL) and saturated brine (50mL).

The organic layer was dried over MgSO₄, filtered and evaporated. Thecrude product was purified by flash silica chromatography, elutiongradient 20 to 100% EtOAc in isohexane. Pure fractions were evaporatedto dryness to afford ethyl4-((tert-butoxycarbonylamino)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylate(468 mg, 53.9%) as a colourless oil which solidified on standing.

1H NMR (400.13 MHz, DMSO-d6) δ 1.22 (3H, t), 1.36-1.38 (9H, m),1.42-1.49 (2H, m), 2.05 (2H, d), 3.13 (2H, d), 3.20 (2H, t), 4.09-4.14(2H, m), 4.45 (2H, d), 6.58 (1H, d), 6.94 (1H, t), 7.16 (1H, d), 8.13(1H, d), 11.65 (1H, s).

MS m/e MH⁺404.

Intermediate 12:4-((tert-butoxycarbonylamino)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid

Lithium hydroxide monohydrate (0.556 g, 13.26 mmol) was added to ethyl4-((tert-butoxycarbonylamino)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylate(Intermediate 11) (1.07 g, 2.65 mmol) in water (6.25 ml), THF (25 ml)and ethanol (25.00 ml). The resulting solution was stirred at 20° C. for1 day. The reaction mixture was diluted with EtOAc (20 mL) and washedwith water (20 mL). The aqueous was adjusted to pH5 with 1M citric acidsolution then extracted with EtOAc (3×50 mL). The organic extracts werewashed with saturated brine (25 mL) then dried over MgSO₄, filtered andevaporated to afford desired product4-((tert-butoxycarbonylamino)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (0.628 g, 63.1%) as a white foam.

1H NMR (400.13 MHz, DMSO-d6) δ 1.36 (9H, s), 1.44-1.51 (2H, m),1.99-2.04 (2H, m), 3.14 (2H, d), 3.25 (2H, s), 4.43-4.46 (2H, m), 6.64(1H, s), 6.84 (1H, t), 7.21 (1H, s), 8.16 (1H, s), 11.82 (1H, s)

MS m/e MH⁺376.

Intermediate 13: (S)-tert-butyl(4-(1-(4-chlorophenyl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-yl)methylcarbamate

HATU (0.251 g, 0.66 mmol) was added in one portion to4-((tert-butoxycarbonylamino)methyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 12) (0.225 g, 0.6 mmol),(S)-1-(4-chlorophenyl)ethanamine (0.093 g, 0.60 mmol) and DIPEA (0.314mL, 1.80 mmol) in DMA (10 mL) at 25° C. under nitrogen. The resultingsolution was stirred at 60° C. for 4 hours. The crude product waspurified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using methanol. Residual HATUwas removed by passing the methanol solution through a silica-supportedcarbonate column. The crude product thus obtained was evaporated todryness to afford (S)-tert-butyl(4-(1-(4-chlorophenyl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-yl)methylcarbamate(0.257 g, 83%) as a colourless gum. This material was used directly inthe next step without further purification.

1H NMR (400.13 MHz, DMSO-d6) δ 1.37 (9H, s), 1.38 (3H, d), 1.48-1.55(2H, m), 2.17 (2H, d), 3.12-3.36 (4H, m), 4.28-4.34 (2H, m), 4.95-5.03(1H, m), 6.65 (2H, s), 7.23-7.24 (1H, m), 7.35 (4H, s), 8.10 (1H, d),8.18 (1H, s), 11.94 (1H, s).

MS m/e MH⁺513.

Intermediate 14: ethyl 4-(4-chlorophenyl)-4-(methoxyimino)butanoate

4-(4-Chlorophenyl)-4-oxobutanoic acid (2.0 g, 9.41 mmol), methoxylaminehydrochloride (0.982 g, 11.76 mmol) and sodium carbonate (0.472 ml,11.29 mmol) in ethanol (30 ml) were stirred and heated at 80° C. for 4hours. The resulting mixture was allowed to cool to room temperature andfiltered. The filtrate was concentrated by evaporation then purified byflash silica chromatography, eluting with 1% TBME in isohexane. Purefractions were evaporated to dryness to afford ethyl4-(4-chlorophenyl)-4-(methoxyimino)butanoate (1.790 g, 70.6%) as acolourless oil.

1H NMR (399.902 MHz, CDCl₃) δ 1.23 (3H, t), 2.53 (2H, t), 3.01 (2H, t),3.98 (3H, s), 4.11 (2H, q), 7.33 (2H, d), 7.58 (2H, d).

MS m/e MH⁺270.

Intermediate 15: 4-(4-chlorophenyl)-4-(methoxyimino)butanoic acid

Lithium hydroxide (0.632 g, 26.40 mmol) was added in one portion at 20°C. to a solution of ethyl 4-(4-chlorophenyl)-4-(methoxyimino)butanoate(Intermediate 14) (1.78 g, 6.60 mmol) in THF (30 ml) and water (20 ml).The resulting solution was stirred at room temperature for 6 hours thenacidified with dilute HCl and extracted with TBME (2×). The combinedextracts were washed with brine, dried over MgSO₄ and evaporated to give4-(4-chlorophenyl)-4-(methoxyimino)butanoic acid (1.520 g, 95%) as acolourless solid.

1H NMR (399.902 MHz, CDCl₃) δ 2.61 (2H, t), 3.01 (2H, t), 3.99 (3H, s),7.34 (2H, d), 7.58 (2H, d).

Intermediate 16: 4-amino-4-(4-chlorophenyl)butan-1-ol

4-(4-Chlorophenyl)-4-(methoxyimino)butanoic acid (Intermediate 15) (6.28g, 26.00 mmol) in tetrahydrofuran (8 ml) was cooled, under an atmosphereof N2, in an ice-methanol bath then treated dropwise withborane-tetrahydrofuran complex (1.0M in THF) (26 ml, 26.00 mmol) over aperiod of 20 minutes. The resulting solution was allowed to warm to roomtemperature, stirred for 1 hour then heated at reflux for a further 6hours. After cooling in ice-water the mixture was treated with water (6ml) dropwise with stirring over 10 minutes. The mixture was againallowed to warm to room temperature and stirred for 2 hours beforeevaporating the bulk of the solvent. The residue was then cooled inice-water and 50% NaOH(aq.) (6 ml) added dropwise with stirring. Theresulting mixture was stirred and heated at 90° C. for 4 hours thencooled to room temperature and extracted with Et₂O (3×). The combinedextracts were washed with water followed by brine, dried over MgSO₄ andevaporated to give 4-amino-4-(4-chlorophenyl)butan-1-ol (1.100 g, 21.2%)as a colourless, viscous oil which was used without furtherpurification.

1H NMR (399.902 MHz, CDCl₃) δ 1.58-1.84 (4H, m), 2.39 (3H, br. s),3.57-3.67 (2H, m), 3.88-3.91 (1H, m), 7.23 (2H, d), 7.30 (2H, d).

MS m/e MH⁺200.

Intermediate 17: tert-butyl4-(1-(4-chlorophenyl)-4-hydroxybutylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (166 mg, 0.44 mmol) was added portionwise to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (150 mg, 0.42 mmol),4-amino-4-(4-chlorophenyl)butan-1-ol (Intermediate 16) (83 mg, 0.42mmol) and N-ethyldiisopropylamine (0.087 ml, 0.50 mmol) in DMF (2.0 mL)at 20° C. The resulting solution was stirred at 20° C. for 3 hours thenquenched in water (10 ml) to give a pale yellow ppt. The precipitate wascollected by filtration, washed with water and dried under vacuum toafford tert-butyl4-(1-(4-chlorophenyl)-4-hydroxybutylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(152 mg, 67.4%) as a cream solid, which was used without furtherpurification.

1H NMR (399.902 MHz, DMSO) δ 1.40 (11H, s), 1.69-1.74 (2H, m), 1.93-2.09(4H, m), 3.38-3.43 (2H, m), 3.51-3.61 (2H, m), 4.19-4.27 (2H, m), 4.38(1H, br s), 4.76 (1H, q), 6.65 (1H, m), 6.95 (1H, s), 7.20 (1H, m), 7.33(4H, s), 7.89 (1H, d), 11.78 (1H, s).

MS m/e MH⁺543.

Intermediate 18: tert-butyl4-(1-(4-chlorophenyl)-2-hydroxyethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (148 mg, 0.39 mmol) was added in one portion to astirred solution of4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (134 mg, 0.37 mmol) and N-ethyldiisopropylamine(0.077 mL, 0.44 mmol) in NMP (3 mL). The mixture was treated with2-amino-2-(4-chlorophenyl)ethanol (70 mg, 0.41 mmol) (CAS™ no.179811-64-4, see US2006/0004045 for preparation). The dark solution wasstirred for 16 hours at room temperature. The mixture was partitionedbetween ethyl acetate and aqueous sodium bicarbonate solution. Theorganic layer was washed twice with water and then brine. The organicsolution was dried over magnesium sulfate, filtered and evaporated. Theresidue was purified by flash silica chromatography on silica usinggradient elution (1% methanol/DCM to 15% methanol/DCM). Productcontaining fractions were combined to give tert-butyl4-(1-(4-chlorophenyl)-2-hydroxyethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(152 mg, 80%) as colourless solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.42 (9H, s), 1.93-2.12 (4H, m), 3.45-3.63(4H, m), 4.22-4.33 (2H, m), 4.75-4.88 (2H, m), 6.59-6.61 (1H, m),7.14-7.24 (2H, m), 7.33 (4H, s), 7.76 (1H, d), 8.14 (1H, s), 11.65 (1H,br, s) m/z (ESI+) (M+H)+=515; HPLC t_(R)=1.99 min.

Intermediate 19: (S)-methyl3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propanoate

Iodomethane (1.038 mL, 16.68 mmol) was added in one portion to(S)-3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propanoic acid (1 g,3.34 mmol) and potassium carbonate (0.922 g, 6.67 mmol) in DMF (15 mL).The resulting suspension was stirred at 80° C. for 24 hours. Thereaction mixture was concentrated and diluted with EtOAc (50 mL) withwater (50 mL). The organic layer was dried over MgSO₄, filtered andevaporated to afford (S)-methyl3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propanoate (1.340 g,128%) as a orange solid. 1H NMR (399.9 MHz, DMSO-d6) δ 1.36 (9H, s),2.71-2.74 (1H, m), 2.74-2.80 (1H, m), 3.57 (3H, s), 4.91 (1H, d), 7.33(2H, d), 7.39 (2H, d), 7.49 (1H, d). m/z (ESI−) (M−H)−=312; HPLC tR=2.57min.

Intermediate 20: (S)-methyl 3-amino-3-(4-chlorophenyl)propanoate(hydrochloride)

Hydrochloric acid (4.0M in 1,4 Dioxane) (4.18 mL, 16.73 mmol) was addedin one portion to (S)-methyl3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propanoate (Intermediate19) (1.05 g, 3.35 mmol) in DCM (20 mL) at 20° C. The resulting solutionwas stirred at 20° C. for 5 hours. The reaction mixture was evaporatedto give (S)-methyl 3-amino-3-(4-chlorophenyl)propanoate (hydrochloride)(0.850 g, 102%) as a white solid. 1H NMR (399.9 MHz, DMSO-d6) δ2.98-3.04 (1H, m), 3.16-3.21 (1H, m), 3.58 (3H, s), 4.62-4.66 (1H, m),7.50-7.52 (2H, m), 7.57-7.59 (2H, m), 8.66 (3H, s). m/z (ESI+)(M+H)+=214; HPLC tR=1.71 min.

Intermediate 21: (S)-methyl3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate

O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluoro-phosphate (1.157 g, 3.04 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (1 g, 2.77 mmol) and N,N-diisopropylethylamine(1.006 mL, 6.09 mmol) in NMP (10 mL) at 20° C. The resulting solutionwas stirred at 20° C. for 5 minutes. (S)-methyl3-amino-3-(4-chlorophenyl)propanoate (hydrochloride) (Intermediate 20)(0.692 g, 2.77 mmol) was then added to the solution and stirred at roomtemperature for 3 hours. The reaction mixture was diluted with EtOAc(100 mL), and washed sequentially with water (2×50 mL) and brine (50mL). The organic layer was dried over MgSO₄, filtered and evaporated toafford crude product. The crude product was purified by flash silicachromatography, elution gradient 0 to 5% MeOH in DCM. Pure fractionswere evaporated to dryness to afford (S)-methyl3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate(1.27 g, 82%) as a white solid. 1H NMR (399.9 MHz, DMSO-d6) δ 1.40 (9H,s), 1.99 (2H, s), 2.04-2.07 (2H, m), 2.79-2.84 (2H, m), 3.56 (3H, s),3.60-3.66 (1H, m), 3.67-3.70 (1H, m), 4.20 (2H, t), 5.20-5.26 (1H, m),6.73 (1H, d), 7.13 (1H, s), 7.27 (1H, t), 7.35 (4H, q), 8.14 (1H, d),8.21 (1H, s), 11.98 (1H, s). m/z (ESI+) (M+H)+=557; HPLC tR=2.12 min.

Intermediate 22: (S)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

LiAlH₄ (2.280 mL, 2.28 mmol) was added dropwise to (S)-methyl3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate(Intermediate 21) (1.27 g, 2.28 mmol) in THF (70 mL) cooled to 0° C.under nitrogen. The resulting solution was stirred at 20° C. for 1 hour.The reaction mixture was quenched with sodium hydroxide (2M) (2 mL) andwater (1 mL). The solution was filtered and was diluted with EtOAc (200mL), and washed sequentially with water (100 mL), water (100 mL), andsaturated brine (100 mL). The organic layer was dried over MgSO₄,filtered and evaporated to afford (S)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(1.04 g, 86%) as a white solid. M/z (ESI+) (M+H)+=529; HPLC tR=2.00 min.

Intermediate 23: 2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)ethylmethanesulfonate

Methanesulfonyl chloride (1.451 mL, 18.74 mmol) was added to tert-butyl1-(4-chlorophenyl)-2-hydroxyethylcarbamate (4.63 g, 17.04 mmol) andN,N-diisopropylethylamine (6.23 mL, 35.78 mmol) in DCM (40 mL) cooled to0° C. over a period of 5 minutes under nitrogen. The resulting solutionwas stirred at 20° C. for 2 hours. The reaction mixture was diluted withDCM (100 mL) and washed sequentially with water (100 mL). The organiclayer was dried over MgSO₄, filtered and evaporated to afford crudeproduct. The crude product was purified by flash silica chromatography,elution gradient 0 to 10% EtOAc in DCM. Pure fractions were evaporatedto dryness to afford2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)ethyl methanesulfonate(3.12 g, 52.3%) as a white solid. 1H NMR (399.9 MHz, DMSO-d6) δ 1.39(9H, s), 3.17 (3H, s), 4.22-4.28 (2H, m), 4.90 (1H, d), 7.40-7.46 (4H,m), 7.68 (1H, d). m/z (ESI+) (M−H)−=348; HPLC tR=2.32 min.

The tert-butyl 1-(4-chlorophenyl)-2-hydroxyethylcarbamate used in theabove reaction was prepared as follows. 2-Amino-2-(4-chlorophenyl)aceticacid (12 g, 64.65 mmol) was stirred in THF (200 mL) and sodiumborohydride (5.82 g, 153.87 mmol) was added in portions to the stirredmixture under nitrogen. A solution of iodine (16.41 g, 64.65 mmol) inTHF (20 mL) was added dropwise maintaining the temperature below 15° C.using an ice bath. The resulting mixture was warmed to room temperatureand stirred at reflux overnight. The reaction was quenched by theaddition of methanol (40 mL). A portion of this solution was removed (50mL) and partitioned between ethyl acetate and water. The organic layerwas concentrated under reduced pressure. The residue was purified byMPLC on silica using gradient elution (0 to 10% methanol/DCM). Thedesired product, 2-amino-2-(4-chlorophenyl)ethanol (1.318 g, 11.88%),was thus isolated as a colourless solid. 1H NMR (399.9 MHz, CDCl₃) δ2.00 (3H, br, s), 3.48-3.58 (1H, m), 3.68-3.76 (1H, m), 4.02-4.08 (1H,m), 7.23-7.39 (4H, m). m/z (ESI−) (M−H)−=284,286; HPLC tR=2.20 min.

The remainder of the solution was treated with triethylamine (18.02 mL,129.31 mmol) and di-tert-butyl dicarbonate (14.11 g, 64.65 mmol). Themixture was stirred for 2 hours at room temperature before beingpartitioned between ethyl acetate and water. The organic layer was driedwith magnesium sulfate, filtered, and concentrated under reducedpressure. The residue was purified by MPLC on silica using gradientelution (10% ethyl acetate/DCM to 50% ethyl acetate/DCM). The desiredproduct, tert-butyl 1-(4-chlorophenyl)-2-hydroxyethylcarbamate (7.91 g,45.0%), was thus isolated as a colourless solid. Impure fractions wererepurified by MPLC to give a second crop (2.41 g, 14%). 1H NMR (399.9MHz, DMSO-d6) δ 1.37 (9H, s), 3.41-3.52 (2H, m), 4.42-4.58 (1H, m), 4.79(1H, t), 7.23 (1H, d), 7.31 (2H, d), 7.37 (2H, d).

Intermediate 24: tert-butyl 1-(4-chlorophenyl)-2-cyanoethylcarbamate

Sodium cyanide (105 mg, 2.14 mmol) was added to2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)ethyl methanesulfonate(Intermediate 23) (300 mg, 0.86 mmol) in DMF (5 mL) at 20° C. Theresulting suspension was stirred at 80° C. for 3 hours. The reactionmixture was evaporated to dryness and redissolved in water (10 mL), andwashed sequentially with DCM 3×(10 mL). The organic layer was dried overMgSO₄, filtered and evaporated to afford crude product. The crudeproduct was purified by flash silica chromatography, elution gradient 0to 25% EtOAc in isohexane. Pure fractions were evaporated to dryness toafford tert-butyl 1-(4-chlorophenyl)-2-cyanoethylcarbamate (209 mg, 87%)as a white solid. 1H NMR (399.9 MHz, DMSO-d6) δ 1.38-1.42 (9H, s),2.82-2.89 (2H, m), 4.89 (1H, d), 7.38-7.45 (4H, m), 7.76 (1H, d). m/z(ESI+) (M−H)−=279; HPLC tR=2.42 min.

Intermediate 25: tert-butyl 3-amino-1-(4-chlorophenyl)propylcarbamate

Lithium aluminium hydride (0.712 mL, 0.71 mmol) was added dropwise totert-butyl 1-(4-chlorophenyl)-2-cyanoethylcarbamate (Intermediate 24)(200 mg, 0.71 mmol) in THF (4 mL) at 20° C. under nitrogen. Theresulting solution was stirred at 20° C. for 2 hours. The reactionmixture was quenched with NaOH (1M) (1 mL) and the solution wasfiltered. The solution was diluted with EtOAc (20 mL), and washed withwater 2×(10 mL). The organic layer was dried over MgSO₄, filtered andevaporated to afford tert-butyl3-amino-1-(4-chlorophenyl)propylcarbamate (203 mg, 100%) as a gum. m/z(ESI+) (M+H)+=285; HPLC tR=2.33 min.

Intermediate 26: tert-butyl3-acetamido-1-(4-chlorophenyl)propylcarbamate

Acetic anhydride (0.084 mL, 0.89 mmol) was added dropwise to tert-butyl3-amino-1-(4-chlorophenyl)propylcarbamate (Intermediate 25) (203 mg,0.71 mmol) and N,N-diisopropylethylamine (0.248 mL, 1.43 mmol) in DCM (5mL) at 20° C. The resulting solution was stirred at 20° C. for 16 hours.The reaction mixture was quenched with NaHCO₃ (2M) (10 mL) and water (10mL) and extracted with DCM (20 mL). The organic layer was dried overMgSO₄, filtered and evaporated to afford crude product. The crudeproduct was purified by flash silica chromatography, elution gradient 0to 2.5% MeOH in DCM. Pure fractions were evaporated to dryness to affordtert-butyl 3-acetamido-1-(4-chlorophenyl)propylcarbamate (142 mg, 61.0%)as a white solid. 1H NMR (399.9 MHz, DMSO-d6) δ 1.36 (9H, s), 1.67-1.82(2H, m), 1.79 (3H, s), 2.97 (2H, q), 4.50 (1H, d), 7.32 (2H, d), 7.38(2H, d), 7.43-7.45 (1H, m), 7.79 (1H, s). m/z (ESI+) (M+H)+=327; HPLCtR=2.03 min.

Intermediate 27: N-(3-amino-3-(4-chlorophenyl)propyl)acetamide

tert-Butyl 3-acetamido-1-(4-chlorophenyl)propylcarbamate (Intermediate26) (142 mg, 0.43 mmol) was added to TFA (2 mL) at 20° C. The resultingsolution was stirred at 20° C. for 40 minutes. The reaction mixture wasevaporated and purified by ion exchange chromatography using an SCXcolumn. The desired product was eluted from the column using 7M NH₃/MeOHand pure fractions were evaporated to dryness to affordN-(3-amino-3-(4-chlorophenyl)propyl)acetamide (39.0 mg, 39.6%) as acolourless gum. m/z (ESI+) (M+H)+=227; HPLC tR=1.32 min.

Intermediate 28: tert-butyl4-(3-acetamido-1-(4-chlorophenyl)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluoro-phosphate (98 mg, 0.26 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (62.2 mg, 0.17 mmol) and N,N-diisopropylethylamine(0.085 mL, 0.52 mmol) in NMP (2 mL) at 20° C. under nitrogen. Theresulting solution was stirred at 20° C. for 5 minutes.N-(3-amino-3-(4-chlorophenyl)propyl)acetamide (Intermediate 27) (39 mg,0.17 mmol) in NMP (2 mL) was then added to the reaction and stirred for1 hour. The reaction mixture was concentrated and diluted with EtOAc (20mL) and washed with water (20 mL). The organic layer was dried overMgSO₄, filtered and evaporated to afford tert-butyl4-(3-acetamido-1-(4-chlorophenyl)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(1.04 g, 86%) as a white solid. m/z (ESI+) (M+H)+=570; HPLC tR=1.95 min.

Intermediate 29: 3-amino-3-(4-chlorophenyl)propan-1-ol

Borane-tetrahydrofuran complex (94.0 mL, 93.92 mmol) was added dropwiseto a stirred suspension of 3-amino-3-(4-chlorophenyl)propionic acid(2.50 g, 12.52 mmol) in THF (75 mL) at 0° C. over a period of 20 minutesunder nitrogen. The resulting suspension was stirred at 0° C. for 30minutes then at 22° C. for 5 hours. The reaction mixture was addedportionwise to methanol (500 mL). The mixture was concentrated,redissolved in methanol (250 mL) and reconcentrated (this process wasrepeated three times). The residue was dissolved in DCM (200 mL) andwashed with 1N NaOH (150 mL). The aqueous layer was extracted with DCM(5×100 mL) and the extracts combined with the organic layer. Thecombined organics were washed with saturated brine (2×150 mL), driedover MgSO₄ and concentrated to afford a white semi-solid. The crudeproduct was purified by flash silica chromatography, elution gradient 5to 7% (10:1 MeOH/conc. NH_(3 (aq))) in DCM. Pure fractions wereevaporated to dryness to afford 3-amino-3-(4-chlorophenyl)propan-1-ol(1.320 g, 56.8%) as a white solid.

1H NMR (399.902 MHz, CDCl₃) δ 1.87 (2H, m), 2.34 (2H, br.s), 3.79 (2H,m), 4.13 (1H, t), 7.24 (2H, d), 7.32 (2H, d).

MS m/e MH⁺169

Intermediate 30: tert-butyl 1-(4-chlorophenyl)-3-hydroxypropylcarbamate

Di-tert-butyl dicarbonate (0.705 g, 3.23 mmol) was added to3-amino-3-(4-chlorophenyl)propan-1-ol (Intermediate 29) (0.500 g, 2.69mmol) in DCM (30 mL) at 22° C. The resulting solution was stirred at 22°C. for 2 hours. The mixture was concentrated and the residue waspurified by flash silica chromatography, elution gradient 0 to 4% (10:1MeOH/conc. NH_(3 (aq))) in DCM. Pure fractions were evaporated todryness to afford tert-butyl 1-(4-chlorophenyl)-3-hydroxypropylcarbamate(0.759 g, 99%) as a white solid.

1H NMR (399.902 MHz, CDCl₃) δ 1.43 (9H, s), 1.81 (1H, m), 2.04 (1H, m),2.74 (1H, br.s), 3.69 (2H, m), 4.88 (1H, br.s), 5.04 (1H, d), 7.23 (2H,d), 7.32 (2H, d).

MS m/e MH⁻284, 286

Intermediate 31: 3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propylmethanesulfonate

Methanesulfonyl chloride (0.097 mL, 1.25 mmol) was added dropwise totert-butyl 1-(4-chlorophenyl)-3-hydroxypropylcarbamate (Intermediate 30)(0.326 g, 1.14 mmol) and triethylamine (0.191 mL, 1.37 mmol) in DCM (15mL) at 22° C. The resulting solution was stirred at 22° C. for 2 hours.The mixture was concentrated and the residue was purified by flashsilica chromatography, elution gradient 20 to 40% EtOAc in isohexane.Pure fractions were evaporated to dryness to afford3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propyl methanesulfonate(0.366 g, 88%) as a white solid.

1H NMR (399.902 MHz, CDCl₃) δ 1.42 (9H, s), 2.19 (2H, m), 3.01 (3H, s),4.24 (2H, m), 4.82 (2H, m), 7.22 (2H, d), 7.33 (2H, d).

MS m/e MH⁻362, 364

Intermediate 32: tert-butyl1-(4-chlorophenyl)-3-(dimethylamino)propylcarbamate

3-(tert-Butoxycarbonylamino)-3-(4-chlorophenyl)propyl methanesulfonate(Intermediate 31) (0.075 g, 0.21 mmol) and tetrabutylammonium iodide(0.015 g, 0.04 mmol) were dissolved in a solution of dimethylamine inTHF (2M, 5.153 mL, 10.31 mmol) and sealed into a microwave tube. Thereaction was heated to 150° C. for 30 minutes in the microwave reactorand cooled to ambient temperature. The reaction mixture wasconcentrated, diluted with DCM (25 mL) and washed with water (25 mL).The organic layer was filtered through a phase-separating filter paperand evaporated. The crude product was purified by flash silicachromatography, elution gradient 4 to 8% (10:1 MeOH/conc. NH_(3 (aq)))in DCM.

Pure fractions were evaporated to dryness to afford tert-butyl1-(4-chlorophenyl)-3-(dimethylamino)propylcarbamate (0.054 mg, 84%) as acolourless oil.

1H NMR (399.902 MHz, CDCl₃) δ 1.40 (9H, s), 1.80 (1H, br.s), 1.94 (1H,m), 2.23 (6H, s), 2.26 (2H, m), 4.71 (1H, br.s), 6.16 (1H, br.s), 7.21(2H, d), 7.29 (2H, d).

MS m/e MH⁺313

Intermediate 33: 1-(4-chlorophenyl)-N3,N3-dimethylpropane-1,3-diamine

Hydrogen chloride (4M in dioxane, 1.132 mL, 32.61 mmol) was added totert-butyl 1-(4-chlorophenyl)-3-(dimethylamino)propylcarbamate(Intermediate 32) (0.051 g, 0.16 mmol) in a mixture of DCM (5 mL) andmethanol (2 mL) at 22° C. The resulting solution was stirred at 22° C.for 4 hours. The mixture was concentrated and the residue was purifiedby ion exchange chromatography, using an SCX column. The desired productwas eluted from the column using 2M NH₃/MeOH and pure fractions wereevaporated to dryness to afford1-(4-chlorophenyl)-N3,N3-dimethylpropane-1,3-diamine (0.032 g, 92%) as acolourless oil.

1H NMR (399.902 MHz, CDCl₃) δ 1.72-1.85 (2H, m), 2.19-2.32 (2H, m), 2.21(6H, s), 3.99 (1H, t), 7.25-7.31 (4H, m).

MS m/e MH⁺213

Intermediate 34: tert-butyl4-(1-(4-chlorophenyl)-3-(dimethylamino)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (0.044 g, 0.12 mmol) was added to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (0.040 g, 0.11 mmol) and N,N-diisopropylethylamine(0.023 mL, 0.13 mmol) in NMP (5 mL) at 22° C. The resulting suspensionwas stirred at 50° C. for 10 minutes. The mixture was cooled to ambienttemperature and 1-(4-chlorophenyl)-N3,N3-dimethylpropane-1,3-diamine(Intermediate 33) (0.023 g, 0.11 mmol) was added as a solution in NMP (2mL). The mixture was stirred at 22° C. for 3 days. The mixture wasdiluted with methanol and purified by ion exchange chromatography usingan SCX column; the desired product was eluted from the column using 30%(2M NH₃ in MeOH) in DCM and product-containing fractions were evaporatedto dryness. The crude product was purified by flash silicachromatography, elution gradient 2 to 6% (10:1 MeOH/conc. NH_(3 (aq)))in DCM. Pure fractions were evaporated to dryness to afford tert-butyl4-(1-(4-chlorophenyl)-3-(dimethylamino)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(0.047 g, 76%) as a white solid.

1H NMR (399.902 MHz, DMSO) δ 1.43 (9H, s), 1.82 (2H, m), 1.90-2.04 (5H,m), 2.14 (6H, s), 2.16 (2H, m), 3.54 (2H, m), 4.25 (2H, m), 4.86 (1H,dt), 6.61 (1H, dd), 7.09 (1H, br.s), 7.17 (1H, dd), 7.33 (4H, s (roofeffect)), 8.14 (1H, s), 8.45 (1H, d), 11.65 (1H, s).

MS m/e MH⁺556

Intermediate 35: methyl3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate

O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluoro-phosphate (84 mg, 0.22 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (72.3 mg, 0.20 mmol),3-amino-3-(4-chloro-phenyl)-propionic acid methyl ester hydrochloride(50 mg, 0.20 mmol) and N,N-diisopropylethylamine (0.104 mL, 0.60 mmol)in N-methyl-2-pyrrolidinone (1.5 mL) at 20° C. under argon. Theresulting solution was stirred for 5 h. The reaction mixture waspurified by preparative HPLC using a Waters X-Bridge reverse-phasecolumn (C-18, 5 microns silica, 19 mm diameter, 100 mm length, flow rateof 40 ml/minute) with decreasingly polar mixtures of water (containing0.2% ammonium carbonate) and acetonitrile as eluent. The fractionscontaining the desired compound were evaporated to dryness to affordmethyl3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate(71.0 mg, 63.8%) as a white crystalline solid.

1H NMR (500 MHz, DMSO-d6) δ 1.36 (9H, s), 1.94-2.00 (4H, m), 2.79-2.86(2H, dq), 3.54 (3H, s), 3.52-3.61 (2H, m), 4.19 (2H, t), 5.22 (1H, q),6.56 (1H, d), 6.9 (1H br s), 7.13 (1H, d), 7.33 (4H, q), 7.98 (1H, d),8.12 (1H, s), 11.53 (1H, s). m/z (ESI+) (M+H)+=557; HPLC tR=3.18 min.

Intermediate 36: tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

Sodium borohydride (1.019 g, 26.93 mmol) was added in one portion to astirred suspension of methyl3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate(Intermediate 35) (1.0 g, 1.80 mmol) dissolved in EtOH (400 mL) underargon. The resulting suspension was stirred at 20° C. for 70 hours. Afew drops of water were added to quench the excess of hydride and themixture stirred for 1 hour. The mixture was evaporated to dryness andthe residue was partitioned between CH₂Cl₂ and water saturated withNaCl. The organic phase was dried and concentrated to provide crudeproduct as white crystals. The crude product was purified by flashchromatography on silica gel eluting with 0 to 10% methanol indichloromethane. The pure fractions were evaporated to dryness to affordtert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(286 mg, 30.1%) as a white solid.

1H NMR (500 MHz, DMSO-d6) δ 1.41 (9H, s), 1.79-1.99 (6H, m), 3.31-3.37(2H, m), 3.51 (2H, m), 4.21-4.28 (2H, m), 4.55 (1H, s), 4.90 (1H, q),6.59 (1H, d), 7.06 (1H, s), 7.16 (1H, d), 7.31 (4H, s), 8.01 (1H, d),8.12 (1H, s), 11.67 (1H, s). m/z (ESI+) (M−H)−=529; HPLC tR=2.93 min.

Intermediate 37: tert-butyl4-(1-(4-chlorophenyl)-3-(1,3-dioxoisoindolin-2-yl)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

Triphenylphosphine (158 mg) and phthalimide (22.11 mg) were added in oneportion to a stirred solution of tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 36) (53 mg) dissolved in THF (10 mL) and cooled to 0° C.under argon. The resulting solution was stirred at 0° C. for 30 minutes.Diethyl azodicarboxylate (0.093 mL) was added dropwise at 0° C. underargon. The resulting suspension was stirred at 0° C. for 60 minutes andovernight at room temperature. The suspension was quenched with water.The THF was removed in vacuo and the aqueous phase extracted with DCM.The organic phase was dried and concentrated to provide crude product.The crude product was purified by flash chromatography on silica geleluting with 0 to 10% methanol in DCM. The solvent was evaporated todryness to afford tert-butyl4-(1-(4-chlorophenyl)-3-(1,3-dioxoisoindolin-2-yl)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamateas a white solid (14.00 mg, 21.23%).

¹H NMR (500 MHz, DMSO-d6) δ 1.37 (9H, s), 2.01 (4H, m), 2.09 (2H, m),3.58 (4H, m), 4.23 (2H, m), 4.84 (1H, q), 6.60 (1H, d), 7.00 (1H, m),7.16 (1H, t), 7.27 (2H, d), 7.33 (2H, d), 7.82 (4H, m), 8.12 (1H, d),8.13 (1H, s), 11.67 (1H, s).

MS m/e MH⁺658.

Intermediate 38: tert-butyl4-(3-amino-1-(4-chlorophenyl)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

Hydrazine monohydrate (10.32 μL) was added to a stirred suspension oftert-butyl4-(1-(4-chlorophenyl)-3-(1,3-dioxoisoindolin-2-yl)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(Intermediate 37) (14 mg) in ethanol (1.0 mL) under argon. The resultingsuspension was stirred at room temperature for 3 days. The mixture wasfiltered and was purified by preparative HPLC using a Waters X-Bridgereverse-phase column (C-18, 5 microns silica, 19 mm diameter, 100 mmlength) and decreasingly polar mixtures of water (containing 0.2%ammonium carbonate) and acetonitrile as eluent. Pure fractions wereevaporated to dryness to afford tert-butyl4-(3-amino-1-(4-chlorophenyl)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamateas a white powder (5.00 mg, 44.5%).

¹H NMR (500 MHz, CDCl₃+CD₃OD) δ 1.43 (9H, s), 1.93 (2H, s), 2.19 (4H,m), 2.74 (2H, s) 3.57 (4H, m), 4.39 (2H, t), 5.01 (1H, t), 6.53 (1H, d),7.07 (1H, d), 7.28 (4H, q), 8.19 (1H, s).

MS m/e MH⁺528.

Intermediate 39: (R)-methyl3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propanoate

Iodomethane (0.987 mL, 15.85 mmol) was added in one portion toboc-(R)-3-amino-3-(4-chloro-phenyl)-propionic acid (950 mg, 3.17 mmol)and potassium carbonate (876 mg, 6.34 mmol) in DMF (15 mL). Theresulting suspension was stirred at 80° C. for 24 hours. The reactionmixture was concentrated and diluted with EtOAc (25 mL) with water (25mL). The organic layer was dried over MgSO₄, filtered and evaporated toafford (R)-methyl3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propanoate (990 mg, 100%)as a orange solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.36 (9H, s), 2.67-2.70 (1H, m), 2.71-2.80(1H, m), 3.57 (3H, s), 4.91 (1H, d), 7.32-7.34 (2H, m), 7.38-7.40 (2H,m), 7.49 (1H, d).

MS m/e MH⁺312; HPLC tR=2.57 min.

Intermediate 40: (R)-methyl 3-amino-3-(4-chlorophenyl)propanoate(hydrochloride)

Hydrochloric acid (4.0M in dioxane) (7.89 mL, 31.55 mmol) was added inone portion to (R)-methyl3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propanoate (Intermediate39) (990 mg, 3.16 mmol) in DCM (20 mL) at 20° C. The resulting solutionwas stirred at 20° C. for 3 hours. The reaction mixture was evaporatedto afford (R)-methyl 3-amino-3-(4-chlorophenyl)propanoate(hydrochloride) (777 mg, 98%) as a yellow solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 2.99-3.05 (1H, m), 3.18-3.23 (1H, m), 3.58(3H, d), 4.62-4.65 (1H, m), 7.49-7.53 (2H, m), 7.57-7.61 (2H, m), 8.72(3H, s).

MS m/e MH⁺214; HPLC tR=1.71 min.

Intermediate 41: (R)-methyl3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate

O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluoro-phosphate (0.579 g, 1.52 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (0.5 g, 1.38 mmol) and DIPEA (0.503 mL, 3.04 mmol)in NMP (5 mL) at 20° C. The resulting solution was stirred at 20° C. for5 minutes. (R)-methyl 3-amino-3-(4-chlorophenyl)propanoate(hydrochloride) (Intermediate 40) (0.346 g, 1.38 mmol) was then added tothe solution and stirred at room temperature for 3 hours. The reactionmixture was diluted with EtOAc (100 mL), and washed sequentially withwater (50 mL), water (50 mL), and water (50 mL). The organic layer wasdried over MgSO₄, filtered and evaporated to afford crude product. Thecrude product was purified by flash silica chromatography, elutiongradient 0 to 5% MeOH in DCM. Pure fractions were evaporated to drynessto afford (R)-methyl3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate(0.800 g, 100%) as a white solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.40 (9H, s), 1.94-1.98 (2H, m), 2.00-2.01(2H, m), 2.83 (1H, d), 2.85-2.87 (2H, m), 3.55 (3H, s), 3.60 (2H, s),4.21 (2H, s), 5.23 (1H, d), 6.60-6.61 (1H, m), 7.16-7.18 (1H, m), 7.35(4H, q), 8.09-8.14 (2H, m), 11.65 (1H, s).

MS m/e MH⁺557; HPLC tR=2.29 min.

Intermediate 42: (R)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

LiAlH₄ (1.398 mL, 1.40 mmol) was added dropwise to (R)-methyl3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate(Intermediate 41) (779 mg, 1.40 mmol) in THF (40 mL) cooled to 0° C.under nitrogen. The resulting solution was stirred at 20° C. for 3hours. The reaction mixture was quenched with water (1 mL) andneutralised with 2M HCl. The reaction mixture was evaporated to drynessand redissolved in DCM (100 mL) and washed with water (100 mL). Theorganic layer was dried over MgSO₄, filtered and evaporated to affordcrude product. The crude product was purified by flash silicachromatography, elution gradient 0 to 10% MeOH in DCM. The purefractions were combined and evaporated to give (R)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(160 mg, 21.63%) as a solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.37-1.41 (9H, s), 1.86-1.92 (2H, m), 1.94(2H, d), 2.00 (2H, d), 3.38 (2H, s), 3.53-3.55 (2H, m), 4.25 (2H, t),4.52 (1H, s), 4.91 (1H, d), 6.60-6.62 (1H, m), 7.01 (1H, s), 7.16-7.18(1H, m), 7.33 (4H, m), 7.98-8.00 (1H, m), 8.14 (1H, s), 11.67 (1H, s).

MS m/e MH⁺529; HPLC tR=2.01 min.

Intermediate 43: 1-(tert-butoxycarbonyl)-4-cyanopiperidine-4-carboxylicacid

An aqueous solution of lithium hydroxide (2M, 39.0 mL, 77.92 mmol) wasadded to a stirred solution of 1-tert-butyl 4-ethyl4-cyanopiperidine-1,4-dicarboxylate (Intermediate 7) (5.5 g, 19.48mmol), in THF (78 mL) at 25° C. The resulting mixture was stirred at 25°C. for 3 hours and monitored by TLC. The reaction mixture was dilutedwith diethyl ether (150 mL), and washed with water (100 mL). The aqueouslayers were combined and then acidified with citric acid (1N, 200 mL).The product was extracted into DCM. The organic layer was dried overmagnesium sulfate, filtered and evaporated to afford1-(tert-butoxycarbonyl)-4-cyanopiperidine-4-carboxylic acid (4.50 g,91%). This material was used in the next steps without furtherpurification.

¹H NMR (400 MHz, CDCl₃) δ 1.40 (9H, s), 1.87-1.97 (2H, m), 2.04 (2H, d),3.08 (2H, t), 4.05 (2H, s), 8.23 (1H, s).

Intermediate 44:(R)—N-(1-(4-chlorophenyl)ethyl)-4-cyanopiperidine-4-carboxamide

O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (1.255 g, 3.30 mmol) was added in one portion to1-(tert-butoxycarbonyl)-4-cyanopiperidine-4-carboxylic acid(Intermediate 43) (0.763 g, 3 mmol), (R)-1-(4-chlorophenyl)ethanamine(0.462 mL, 3.00 mmol) and DIPEA (1.572 mL, 9.00 mmol) in DMA (20 mL) at25° C. under nitrogen. The resulting solution was stirred at 60° C. for4 hours. The reaction mixture was evaporated to dryness and redissolvedin DCM (150 mL), and washed sequentially with 1M aqueous citric acid (50mL), water (50 mL), and saturated sodium hydrogen carbonate (100 mL).The organic layer was dried over magnesium sulfate, filtered andevaporated to afford crude product. The crude product thus obtained wasconcentrated, then purified by flash silica chromatography, elutiongradient 0 to 100% EtOAc in isohexane. Pure fractions were evaporated toafford (R)-tert-butyl4-(1-(4-chlorophenyl)ethylcarbamoyl)-4-cyanopiperidine-1-carboxylate(1.100 g, 94%) as a colourless gum which solidified on drying under highvacuum. This material was then redissolved in DCM (20.00 mL), andtrifluoroacetic acid (2.311 mL, 30.00 mmol) was added. The solution wasstirred at room temperature for 3 hours, after which the crude productwas purified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using 7N ammonia/MeOH andpure fractions were evaporated to dryness to afford(R)—N-(1-(4-chlorophenyl)ethyl)-4-cyanopiperidine-4-carboxamide (0.720g, 82%) as a colourless gum. This material was used directly in the nextstep without further purification.

MS m/e MH⁺292.

Intermediate 45:(R)—N-(1-(4-chlorophenyl)ethyl)-4-cyano-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

DIPEA (1.101 mL, 6.17 mmol) was added to(R)—N-(1-(4-chlorophenyl)ethyl)-4-cyanopiperidine-4-carboxamide(Intermediate 44) (720 mg, 2.47 mmol) and 6-chloro-7-deazapurine (379mg, 2.47 mmol) in DMA (50 mL) at 25° C. The resulting solution wasstirred at 90° C. for 3 hours. The crude product was purified by ionexchange chromatography, using an SCX column. The desired product waseluted from the column using 7N ammonia/MeOH and pure fractions wereevaporated to dryness to afford(R)—N-(1-(4-chlorophenyl)ethyl)-4-cyano-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(46.2%) as an orange solid. This was used in the next step withoutfurther purification.

¹H NMR (400 MHz, DMSO) δ 1.38 (3H, d), 1.94-2.09 (3H, m), 2.19 (3H, t),4.72 (2H, d), 4.92 (1H, dd), 6.61-6.72 (1H, m), 7.23 (1H, dd), 7.30-7.41(4H, m), 8.18 (1H, s), 8.77 (1H, d), 11.77 (1H, s).

MS m/e MH⁺409.

Intermediate 46: (S)-4-(1-amino-3-hydroxypropyl)benzonitrile

Sodium borohydride (1.039 mL, 29.48 mmol) was added portionwise to(S)-methyl 3-amino-3-(4-cyanophenyl)propanoate (2.23 g, 10.92 mmol) inmethanol (20 mL) at 0° C. over a period of 5 minutes. The resultingsolution was stirred at 20° C. for 24 hours. The reaction mixture wasquenched with saturated NaHCO₃ (50 mL), extracted with EtOAc (3×100 mL),the organic layer was washed with saturated brine (75 mL), dried overMgSO₄, filtered and evaporated to afford crude product. The crudeproduct was purified by flash silica chromatography, elution gradient 0to 8% MeOH with ammonia in DCM. Pure fractions were evaporated todryness to afford (S)-4-(1-amino-3-hydroxypropyl)benzonitrile (0.368 g,19.13%) as a colourless oil.

¹H NMR (400.13 MHz, CDCl₃) δ 1.84-1.93 (2H, m), 3.77-3.80 (2H, m),4.22-4.25 (1H, m), 7.43 (2H, m), 7.63-7.66 (2H, m).

MS m/e MH⁺177.

Intermediate 47: (S)-3-amino-3-(4-chlorophenyl)propan-1-ol

Borane-tetrahydrofuran complex (376 mL, 375.69 mmol) was added dropwiseto a stirred suspension of (S)-3-amino-3-(4-chlorophenyl)propanoic acid(10 g, 50.09 mmol) in THF (500 mL) at 0° C. over a period of 45 minutesunder nitrogen. The resulting suspension was stirred at 0° C. for 30minutes then at 22° C. for 5 hours. The reaction mixture was addedportionwise to methanol (500 mL). The solution was stirred at roomtemperature for three days. The mixture was concentrated, redissolved inmethanol (250 mL) and reconcentrated (this process was repeated threetimes). The residue was dissolved in DCM (75 mL) and washed with 1N NaOH(50 mL). The aqueous layer was extracted with DCM (3×100 mL) and theextracts combined with the organic layer. The combined organics werewashed with saturated brine (50 mL), dried over MgSO₄ and concentratedto afford crude product. The crude product was purified by flash silicachromatography, elution gradient 2 to 6% MeOH/ammonia in DCM. Purefractions were evaporated to dryness to afford(S)-3-amino-3-(4-chlorophenyl)propan-1-ol (5.76 g, 61.9%) as a whitesolid.

¹H NMR (400.13 MHz, CDCl₃) δ 1.84-1.93 (2H, m), 3.76-3.81 (2H, m), 4.13(1H, t), 7.23-7.25 (2H, m), 7.30-7.34 (2H, m).

MS m/e MH⁺186.

Intermediate 48: (S)-tert-butyl4-(tert-butoxycarbonylamino)-4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)piperidine-1-carboxylate

(S)-3-amino-3-(4-chlorophenyl)propan-1-ol (Intermediate 47) (1.09 g,5.87 mmol) was added in one portion to1-(tert-butoxycarbonyl)-4-(tert-butoxycarbonylamino)piperidine-4-carboxylicacid (Intermediate 70) (2.022 g, 5.87 mmol) and DIPEA (3.08 mL, 17.61mmol) in DMA (20 mL).O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (2.456 g, 6.46 mmol) was added and the resultingsolution was stirred at 20° C. for 24 hours. The reaction mixture wasevaporated to dryness then diluted with EtOAc (300 mL), and washedsequentially with water (50 mL) and saturated brine (50 mL). The organiclayer was dried over MgSO₄, filtered and evaporated then triturated withdiethyl ether to afford crude (S)-tert-butyl4-(tert-butoxycarbonylamino)-4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)piperidine-1-carboxylate(4.66 g, 155%) as a white solid.

1H NMR (400.13 MHz, DMSO-d6) δ 1.39 (18H, s), 1.71-1.92 (6H, m), 3.06(2H, s), 3.36 (2H, t), 3.54-3.65 (2H, m), 4.52 (1H, t), 4.89 (1H, q),6.89 (1H, s), 7.29-7.34 (4H, m).

MS m/e M+Na⁺534.

Intermediate 49:(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide

Hydrogen chloride 4M in dioxane (11.72 mL, 46.87 mmol) was added to(S)-tert-butyl4-(tert-butoxycarbonylamino)-4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)piperidine-1-carboxylate(Intermediate 48) (3 g, 5.86 mmol) in dioxane (30 mL). The resultingsolution was stirred at 20° C. for 2 hours. The reaction mixture wasdissolved in methanol and purified by ion exchange chromatography, usingan SCX column. The desired product was eluted from the column using 3.5Nammonia/MeOH and pure fractions were evaporated to dryness to afford(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide(1.970 g, 108%) as a colourless gum.

¹H NMR (400.13 MHz, DMSO-d6) δ 1.20-1.28 (2H, m), 1.75-1.91 (4H, m),2.67-2.83 (2H, m), 3.30 (2H, m), 4.87 (1H, s), 7.30-7.37 (4H, m).

MS m/e MH⁺312.

Intermediate 50: 5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine

N-Bromosuccinimide (6.84 g, 38.42 mmol) was added portionwise to4-chloro-7H-pyrrolo[2,3-d]pyrimidine (5 g, 32.56 mmol) in DCM, dry (125mL) at 20° C. under nitrogen. The resulting suspension was stirred at20° C. for 1 hour. The reaction mixture was evaporated and the resultingbrown solid was triturated with water to give a purple solid which wascollected by filtration. The crude solid was triturated with hot MeOH togive a solid which was collected by filtration. The hot trituration wasrepeated to give 5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (5.23 g,69.1%) as a cream solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 7.94 (1H, s), 8.63 (1H, s), 12.95 (1H, s)

MS m/e MH⁺234.

Intermediate 51: (S)-3-amino-3-(4-bromophenyl)propan-1-ol

Borane-tetrahydrofuran complex (71.7 mL, 71.70 mmol) was added dropwiseto a stirred suspension of (S)-3-amino-3-(4-bromophenyl)propanoic acid(3.5 g, 14.34 mmol) in THF (80 mL) at 0° C. over a period of 30 minutesunder nitrogen. The resulting suspension was stirred at 0° C. for 30minutes then at 22° C. for 5 hours. The reaction mixture was addedportionwise to methanol (250 mL). The solution was stirred at roomtemperature for 24 hours. The mixture was concentrated, redissolved inmethanol (250 mL) and reconcentrated (this process was repeated threetimes). The residue was dissolved in DCM (75 mL) and washed with 1N NaOH(50 mL). The aqueous layer was extracted with DCM (3×100 mL) and theextracts combined with the organic layer. The combined organics werewashed with saturated brine (50 mL), dried over MgSO₄ and concentratedto afford crude product. The crude product was purified by flash silicachromatography, elution gradient 2 to 6% MeOH/ammonia in DCM. Purefractions were evaporated to dryness to afford(S)-3-amino-3-(4-bromophenyl)propan-1-ol (2.160 g, 65.5%) as acolourless gum.

¹H NMR (400.13 MHz, CDCl₃) δ 1.82-1.90 (2H, m), 3.76-3.82 (2H, m), 4.12(1H, t), 7.17-7.20 (2H, m), 7.45-7.49 (2H, m).

MS m/e MH⁺230.

Intermediate 52: ethyl 4-(4-chlorophenyl)-4-(methoxyimino)butanoate

4-(4-Chlorophenyl)-4-oxobutanoic acid (10 g, 47.03 mmol), methoxylaminehydrochloride (4.91 g, 58.79 mmol) and sodium carbonate (5.98 g, 56.44mmol) in ethanol (150 mL) were stirred for 4 hrs at 80° C. The mixturewas then cooled to room temperature before being filtered. The filtratewas concentrated by evaporation and purified by flash silicachromatography (eluent 20-50% EtOAc/iso-hexane). Pure fractions wereevaporated to dryness to afford ethyl4-(4-chlorophenyl)-4-(methoxyimino)butanoate (12.33 g, 97%) as an orangetransparent liquid.

¹H NMR (400.13 MHz, DMSO) 1.13-1.17 (3H, t), 2.26-2.50 (2H, t),2.50-2.52 (1H, qu), 2.94-2.98 (2H, t), 3.94 (3H, s), 4.00-4.05 (2H, q),7.47-7.49 (2H, d), 7.66-7.69 (2H, d).

Intermediate 53: 4-(4-chlorophenyl)-4-(methoxyimino)butanoic acid

Lithium hydroxide monohydrate (9.59 g, 228.57 mmol) was added to ethyl4-(4-chlorophenyl)-4-(methoxyimino)butanoate (Intermediate 52) (12.33 g,45.71 mmol) in water (25.4 mL), THF (102 mL) and ethanol (102 mL). Theresulting solution was stirred at 20° C. for 1 day. The reaction mixturewas diluted with EtOAc (200 mL) and washed with water (200 mL). Theaqueous was adjusted to pH5 with 1M citric acid solution then extractedwith EtOAc (3×150 mL). The organic extracts were washed with saturatedbrine (25 mL) then dried over MgSO₄, filtered and evaporated to afforddesired product 4-(4-chlorophenyl)-4-(methoxyimino)butanoic acid (7.63g, 69.1%) as a light yellow solid.

¹H NMR (400.13 MHz) 2.39-2.43 (2H, t), 2.91-2.95 (2H, t), 3.94 (3H, s),7.47-7.49 (2H, d), 7.67-7.69 (2H, d), 12.21 (1H, broad).

MS m/e MH⁺242.

Intermediate 54: 4-amino-4-(4-chlorophenyl)butan-1-ol

4-(4-Chlorophenyl)-4-(methoxyimino)butanoic acid (Intermediate 53) (6.43g, 26.61 mmol) in tetrahydrofuran (30 mL) was cooled, under anatmosphere of nitrogen, in an ice-methanol bath. Borane-tetrahydrofurancomplex (1.0M in THF) (93 mL, 93.12 mmol) was added dropwise over aperiod of 20 minutes. The resulting solution was allowed to warm to roomtemperature, stirred for 1 hour then heated at reflux for a further 6hours. After cooling in ice-water the mixture was treated with water (20mL) dropwise with stirring over 10 minutes. The mixture was againallowed to warm to room temperature and stirred for 2 hours beforeevaporating the bulk of the solvent. The residue was then cooled inice-water and 50% NaOH (aq.) (20 mL) added dropwise with stirring. Theresulting mixture was stirred and heated at 90° C. for 4 hours thencooled to room temperature and extracted three times with Et₂O. Thecombined extracts were washed with water followed by brine, dried overMgSO₄ and evaporated to dryness. The crude product was then purified byflash silica chromatography (eluent 0-10% 7N ammonia in MeOH/DCM) andpure fractions evaporated to afford 4-amino-4-(4-chlorophenyl)butan-1-ol(3.88 g, 73.0%) as a colourless, transparent gum.

¹H NMR (400.13 MHz) 1.25-1.47 (2H, dm), 1.51-1.61 (2H, m), 3.34-3.38(2H, t), 3.76-3.79 (1H, t), 7.33-7.38 (4H, m).

Intermediate 55: tert-butyl 1-(4-chlorophenyl)-4-hydroxybutylcarbamate

Di-tert-butyl dicarbonate (1.588 mL, 6.91 mmol) was added in one portionto 4-amino-4-(4-chlorophenyl)butan-1-ol (Intermediate 54) (1.38 g, 6.91mmol) in DCM (20 mL) at 25° C. under nitrogen. The resulting solutionwas stirred at room temperature for 24 hours. The resulting mixture wasevaporated to dryness and the crude oil was triturated with isohexane togive a solid which was collected by filtration and dried under vacuum togive tert-butyl 1-(4-chlorophenyl)-4-hydroxybutylcarbamate (1.800 g,87%) as a white solid.

¹H NMR (400.13 MHz, CDCl₃) δ 1.40 (9H, s), 1.48-1.66 (2H, m), 1.78-1.83(2H, m), 3.63-3.68 (2H, m), 4.61 (1H, s), 4.84 (1H, s), 7.21 (2H, m),7.29-7.31 (2H, m).

Intermediate 56: 4-(tert-butoxycarbonylamino)-4-(4-chlorophenyl)butylmethanesulfonate

Methanesulfonyl chloride (0.511 mL, 6.60 mmol) was added dropwise totert-butyl 1-(4-chlorophenyl)-4-hydroxybutylcarbamate (Intermediate 55)(1.80 g, 6.00 mmol) and triethylamine (1.004 mL, 7.20 mmol) in DCM (30.0mL) at 25° C. over a period of 15 minutes under nitrogen. The resultingsolution was stirred at room temperature for 30 minutes. The resultingmixture was evaporated to dryness and the residue was suspended in DCM,filtered and the filtrate purified by flash silica chromatography,elution gradient 20 to 100% EtOAc in isohexane. Pure fractions wereevaporated to dryness to afford4-(tert-butoxycarbonylamino)-4-(4-chlorophenyl)butyl methanesulfonate(2.270 g, 100%) as a white solid.

¹H NMR (400.13 MHz, CDCl₃) δ 1.41 (9H, s), 1.67-1.87 (4H, m), 2.99 (3H,s), 4.23 (2H, t), 4.62 (1H, s), 4.75 (1H, s), 7.20 (2H, d), 7.30-7.33(2H, d).

MS m/e M+Na⁺400.

Intermediate 57: 1-(4-chlorophenyl)-N4,N4-dimethylbutane-1,4-diamine

Dimethylamine (84 mg, 1.85 mmol) and4-(tert-butoxycarbonylamino)-4-(4-chlorophenyl)butyl methanesulfonate(Intermediate 56) (700 mg, 1.85 mmol) were dissolved in THF (10 mL) andsealed into a microwave tube. The reaction was heated to 120° C. for 40minutes in the microwave reactor and cooled to room temperature. Thereaction mixture was concentrated and then dissolved in DCM (10.00 mL)and TFA (2 mL). The reaction was stirred at 20° C. for 2 hours. Thecrude product was purified by ion exchange chromatography, using an SCXcolumn. The desired product was eluted from the column using 3.5Nammonia/MeOH and pure fractions were evaporated to dryness to afford1-(4-chlorophenyl)-N4,N4-dimethylbutane-1,4-diamine (366 mg, 87%) as ayellow gum.

¹H NMR (400.13 MHz, CDCl₃) δ 0.70-0.79 (1H, m), 0.82-0.91 (1H, m),0.93-1.09 (2H, m), 1.21 (6H, s), 1.64 (2H, t), 3.23 (1H, t), 6.58-6.66(4H, m).

MS m/e MH⁺227.

Intermediate 58: (S)-tert-butyl1-(4-chlorophenyl)-3-hydroxypropylcarbamate

Di-tert-butyl dicarbonate (8.04 mL, 35.01 mmol) was added in one portionto (S)-3-amino-3-(4-chlorophenyl)propan-1-ol (Intermediate 47) (6.5 g,35.01 mmol) in DCM (200 mL) at 25° C. under nitrogen. The resultingsolution was stirred at room temperature for 24 hours. The resultingmixture was evaporated to dryness and the crude oil was triturated withisohexane to give a solid which was collected by filtration and driedunder vacuum to give (5)-tert-butyl1-(4-chlorophenyl)-3-hydroxypropylcarbamate (9.20 g, 92%) as a whitesolid.

¹H NMR (399.9 MHz, CDCl₃) δ 1.43 (9H, s), 1.78-1.84 (1H, m), 2.05 (1H,d), 2.74 (1H, s), 3.67-3.71 (2H, m), 4.88 (1H, s), 5.04 (1H, d),7.21-7.24 (2H, m), 7.30-7.33 (2H, m).

MS m/e M+Na⁺308.

Intermediate 59:(S)-3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propylmethanesulfonate

Methanesulfonyl chloride (2.74 mL, 35.41 mmol) was added dropwise to(S)-tert-butyl 1-(4-chlorophenyl)-3-hydroxypropylcarbamate (Intermediate58) (9.2 g, 32.19 mmol) and triethylamine (5.38 mL, 38.63 mmol) in DCM(161 mL) at 25° C. over a period of 15 minutes under nitrogen. Theresulting solution was stirred at room temperature for 30 minutes. Theresulting mixture was evaporated to dryness and the residue was purifiedby flash silica chromatography, elution gradient 40 to 100% EtOAc inisohexane. Pure fractions were evaporated to dryness to afford(S)-3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propylmethanesulfonate (10.03 g, 86%) as a white solid.

¹H NMR (399.9 MHz, CDCl₃) δ 1.36-1.48 (9H, m), 2.20 (2H, s), 3.01 (3H,s), 4.19-4.29 (2H, m), 4.81 (2H, s), 7.21-7.23 (2H, d), 7.32-7.35 (2H,d).

MS m/e M+Na⁺386.

Intermediate 60: (S)-1-(4-chlorophenyl)-N3,N3-diethylpropane-1,3-diamine

Diethylamine (0.426 mL, 4.12 mmol) and(S)-3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propylmethanesulfonate (Intermediate 59) (500 mg, 1.37 mmol) were dissolved inTHF (10 mL) and sealed into a microwave tube. The reaction was heated to120° C. for 50 minutes in the microwave reactor and cooled to roomtemperature. The reaction mixture was concentrated and diluted with DCM(5 mL). TFA (1 mL) was added and the reaction was stirred at 20° C. for1 hour. The crude product was purified by ion exchange chromatography,using an SCX column. The desired product was eluted from the columnusing 3.5N ammonia/MeOH and pure fractions were evaporated to dryness toafford (S)-1-(4-chlorophenyl)-N3,N3-diethylpropane-1,3-diamine (119 mg,36.0%) as a yellow oil.

¹H NMR (400.13 MHz, CDCl₃) δ 1.00 (6H, t), 1.75-1.82 (2H, m), 2.40-2.54(6H, m), 3.97 (1H, t), 7.27-7.31 (4H, m).

MS m/e MH⁺241.

Intermediate 61: 5-bromo-4-chloro-7-tosyl-7H-pyrrolo[2,3-d]pyrimidine

5-Bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (Intermediate 50) (2 g,8.60 mmol) and p-toluenesulfonyl chloride (1.640 g, 8.60 mmol) was takenup in acetone (12 mL) and stirred under nitrogen and cooled to between−5° C. and 5° C. To this solution was added 2.0M NaOH solution (5.50 mL,10.99 mmol) while maintaining the internal temperature at less than 5°C. The reaction was allowed to warm to room temperature and stirred for1 hour. The white solid was filtered off and washed well with acetone toafford 5-bromo-4-chloro-7-tosyl-7H-pyrrolo[2,3-d]pyrimidine (3.28 g,99%) as a pale cream solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 2.38 (3H, s), 7.48-7.50 (2H, m),8.06-8.08 (2H, m), 8.41 (1H, s), 8.84 (1H, s).

MS m/e MH⁺387.

Intermediate 62: methyl1-(5-bromo-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-4-(tert-butoxycarbonylamino)piperidine-4-carboxylate

Triethylamine (3.55 mL, 25.45 mmol) was added to5-bromo-4-chloro-7-tosyl-7H-pyrrolo[2,3-d]pyrimidine (Intermediate 61)(3.28 g, 8.48 mmol) and methyl4-(tert-butoxycarbonylamino)piperidine-4-carboxylate (WO2008075109)(3.07 g, 11.88 mmol) in DMA (50 mL) at 20° C. The resulting suspensionwas stirred at 70° C. for 2 hours. The reaction mixture was concentratedand diluted with EtOAc (200 mL), and washed sequentially with water (75mL) and saturated brine (75 mL). The organic layer was dried over MgSO₄,filtered and evaporated to afford crude product. The crude product waspurified by flash silica chromatography, elution gradient 30 to 70%EtOAc in isohexane. Pure fractions were evaporated to dryness to affordmethyl1-(5-bromo-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-4-(tert-butoxycarbonylamino)piperidine-4-carboxylate(3.60 g, 69.7%) as a white solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 1.38 (9H, s), 2.03 (2H, s), 2.38 (3H, s),3.33-3.37 (2H, m), 3.59 (3H, s), 3.84 (2H, d), 7.41 (1H, s), 7.46 (2H,d), 8.00 (1H, s), 8.03 (2H, d), 8.40 (1H, s).

MS m/e MH⁺610.

Intermediate 63: methyl4-(tert-butoxycarbonylamino)-1-(5-cyclopropyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylate

Methyl1-(5-bromo-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-4-(tert-butoxycarbonylamino)piperidine-4-carboxylate(Intermediate 62) (1.1 g, 1.81 mmol), potassium phosphate tribasic(1.343 g, 6.33 mmol), tricyclohexyl phosphine (0.101 g, 0.36 mmol) andcyclopropylboronic acid (0.438 g, 4.34 mmol) were taken up in toluene(10 mL) and water (0.400 mL). The mixture was purged with nitrogen for30 minutes then palladium(II) acetate (0.041 g, 0.18 mmol) was added.The mixture was heated to 75° C. for 6 hours and then allowed to cooland stir at room temperature overnight. The reaction mixture was dilutedwith EtOAc (75 mL), and washed sequentially with water (50 mL) andsaturated brine (50 mL). The organic layer was dried over MgSO₄,filtered and evaporated to afford crude product. The crude product waspurified by flash silica chromatography, elution gradient 10 to 30%EtOAc in isohexane. Pure fractions were evaporated to dryness to affordmethyl4-(tert-butoxycarbonylamino)-1-(5-cyclopropyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylate(0.447 g, 43.4%) as a cream solid.

¹H NMR (400.13 MHz, CDCl₃) δ 0.72-0.76 (2H, m), 0.99-1.04 (2H, m), 1.43(9H, s), 1.90-1.94 (1H, m), 2.04-2.10 (2H, m), 2.17-2.25 (2H, m), 2.38(3H, s), 3.34-3.41 (2H, m), 3.74 (3H, s), 3.98-4.03 (2H, m), 4.76 (1H,s), 7.12 (1H, s), 7.28 (2H, m), 8.03-8.05 (2H, m), 8.46 (1H, s).

MS m/e MH⁺570.

Intermediate 64:4-(tert-butoxycarbonylamino)-1-(5-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid

2M sodium hydroxide solution (5.00 mL) was added to methyl4-(tert-butoxycarbonylamino)-1-(5-cyclopropyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylate(Intermediate 63) (780 mg, 1.37 mmol) in THF (10 mL) at 20° C. Theresulting solution was stirred at 40° C. for 24 hours. The reactionmixture was diluted with EtOAc (50 mL) and the aqueous layer wascollected and acidified to pH4 with 2M HCl solution. The aqueous layerwas extracted with DCM (2×50 mL) and the organic layers combined andwashed with saturated brine (50 mL). The organic layer was dried overMgSO₄, filtered and evaporated to afford crude product4-(tert-butoxycarbonylamino)-1-(5-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (375 mg, 68.2%) as a cream solid.

¹H NMR (400.13 MHz, DMSO-d6) δ 0.66-0.73 (2H, m), 0.88-1.00 (2H, m),1.40 (9H, s), 1.94-2.09 (5H, m), 3.41-3.53 (2H, m), 3.92-4.01 (2H, m),6.92 (1H, d), 7.18-7.22 (1H, m), 8.21 (1H, s), 11.51 (1H, s), 12.29 (1H,s).

MS m/e MH⁺402.

Intermediate 65: (S)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(5-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

(S)-3-amino-3-(4-chlorophenyl)propan-1-ol (173 mg, 0.93 mmol) was addedin one portion to4-(tert-butoxycarbonylamino)-1-(5-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 64) (375 mg, 0.93 mmol) and DIPEA (0.489 mL, 2.80mmol) in DMA (5 mL).O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (391 mg, 1.03 mmol) was added and the resultingsolution was stirred at 20° C. for 24 hours. The reaction mixture wasevaporated to dryness then diluted with EtOAc (75 mL), and washedsequentially with water (50 mL) and saturated brine (50 mL). The organiclayer was dried over MgSO₄, filtered and evaporated to afford crudeproduct. The crude product was purified by flash silica chromatography,elution gradient 2 to 6% MeOH with ammonia in DCM. Fractions wereevaporated to afford (S)-tert-butyl4-(1-(4-chlorophenyl)-3-hydroxypropylcarbamoyl)-1-(5-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(420 mg, 79%) as a yellow gum.

¹H NMR (400.13 MHz, DMSO-d6) δ 0.66-0.70 (2H, m), 0.86-0.91 (2H, m),1.40 (9H, s), 1.81-2.06 (7H, m), 3.33-3.40 (4H, m), 3.90-3.94 (2H, m),4.53 (1H, t), 4.91-4.93 (1H, m), 6.92 (1H, s), 7.30-7.38 (4H, m), 7.96(1H, d), 8.20 (1H, s), 11.49 (1H, s).

MS m/e MH⁺569.

Intermediate 66: tert-butyl1-(4-chlorophenyl)-3-(methylamino)propylcarbamate

Methylamine gas was bubbled into a solution of3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propyl methanesulfonate(Intermediate 31) (70 mg, 0.19 mmol) in THF (5 mL) at 22° C. over aperiod of 5 minutes. The mixture was sealed into a microwave tube. Thereaction was heated to 125° C. for 30 minutes in the microwave reactorand cooled to ambient temperature. The mixture was evaporated and theresidue was purified by flash silica chromatography, elution gradient 4to 8% (10:1 MeOH/conc. NH_(3 (aq))) in DCM. Pure fractions wereevaporated to dryness to afford tert-butyl1-(4-chlorophenyl)-3-(methylamino)propylcarbamate (52 mg, 90%) as acolourless oil.

¹H NMR (399.902 MHz, CDCl₃) δ 1.41 (9H, s), 1.80 (1H, br.m), 1.93 (1H,br.m), 2.40 (3H, s), 2.59 (2H, m), 4.73 (1H, br.s), 5.94 (1H, br.s),7.21 (2H, d), 7.29 (2H, d).

MS m/e MH⁺299.5.

Intermediate 67: 1-(4-chlorophenyl)-N3-methylpropane-1,3-diamine

Hydrogen chloride (4M in 1,4-dioxane, 0.50 mL, 2.00 mmol) was added totert-butyl 1-(4-chlorophenyl)-3-(methylamino)propylcarbamate(Intermediate 66) (50 mg, 0.17 mmol) in a mixture of DCM (5 mL) andmethanol (2 mL) at 22° C. The resulting solution was stirred at 22° C.for 4 hours. The mixture was concentrated and the residue was purifiedby ion exchange chromatography, using an SCX column. The desired productwas eluted from the column using 2M NH₃/MeOH and pure fractions wereevaporated to dryness to afford1-(4-chlorophenyl)-N3-methylpropane-1,3-diamine (30 mg, 90%) as acolourless oil.

¹H NMR (399.902 MHz, CDCl₃) δ 1.81 (2H, dt), 2.40 (3H, s), 2.58 (2H, m),4.01 (1H, t), 7.25-7.31 (4H, m).

MS m/e MH⁺199.3.

Intermediate 68: tert-butyl4-(1-(4-chlorophenyl)-3-(methylamino)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (44 mg, 0.12 mmol) was added to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (40 mg, 0.11 mmol) and N,N-diisopropylethylamine(0.023 mL, 0.13 mmol) in NMP (5 mL) at 22° C. The resulting suspensionwas stirred at 50° C. for 10 minutes. The mixture was cooled to ambienttemperature and 1-(4-chlorophenyl)-N3-methylpropane-1,3-diamine(Intermediate 67) (22 mg, 0.11 mmol) was added as a solution in NMP (2mL) and the mixture stirred at 22° C. for 3 days. The reaction mixturewas diluted with EtOAc (50 mL) and washed with water (50 mL). Theaqueous layer was passed through an SCX column. The column was elutedwith water and methanol to remove impurities followed by 30% (2M NH₃ inmethanol) in DCM to elute the product. The appropriate fractions wereevaporated to dryness to afford tert-butyl4-(1-(4-chlorophenyl)-3-(methylamino)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(10.00 mg, 17%) as a white solid.

MS m/e MH⁺542.5.

Intermediate 69: 1-tert-butyl 4-methyl4-(tert-butoxycarbonylamino)piperidine-1,4-dicarboxylate

Methyl 4-(tert-butoxycarbonylamino)piperidine-4-carboxylate(WO2008075109) (10 g, 38.71 mmol) was dissolved in DCM (194 mL). To thiswas added di-tert-butyl dicarbonate (10.67 mL, 46.46 mmol) portionwise.After the addition the reaction was stirred at 25° C. for 1 hour. Thecrude product was washed with saturated sodium bicarbonate (3×50 mL) andthe organic layers dried over MgSO₄ before being evaporated to drynessto afford 1-tert-butyl 4-methyl4-(tert-butoxycarbonylamino)piperidine-1,4-dicarboxylate (13.6 g, 98%)as a white solid.

¹H NMR (400.13 MHz, DMSO) δ 1.39 (9H, s), 1.40 (9H, s), 1.70-1.77 (2H,td), 1.88-1.92 (2H, d), 3.05 (2H, broad), 3.61 (3H, s), 3.62-3.65 (2H,m), 7.33 (broad, exchange).

Intermediate 70:1-(tert-butoxycarbonyl)-4-(tert-butoxycarbonylamino)piperidine-4-carboxylicacid

Lithium hydroxide monohydrate (8.13 g, 193.62 mmol) was added to1-tert-butyl 4-methyl4-(tert-butoxycarbonylamino)piperidine-1,4-dicarboxylate (Intermediate69) (13.88 g, 38.72 mmol) in water (21.51 mL), THF (86 mL) and methanol(86 mL). The resulting solution was stirred at 20° C. for 1 day. Thereaction mixture was diluted with EtOAc (75 mL) and washed with water(75 mL). The aqueous was adjusted to pH5 with 1M citric acid solution,then extracted with EtOAc (3×200 mL). The organic extracts were washedwith saturated brine (50 mL), then dried over MgSO₄, filtered andevaporated to afford desired product1-(tert-butoxycarbonyl)-4-(tert-butoxycarbonylamino)piperidine-4-carboxylicacid (10.36 g, 78%) as a fine white solid.

¹H NMR (400.13 MHz, DMSO) δ 1.39 (9H, s), 1.40 (9H, s), 1.68-1.76 (2H,td), 1.89-1.92 (2H, d), 3.03 (2H, broad), 3.62-3.65 (2H, d), 7.16(exchange), 12.36 (exchange).

Intermediate 71: tert-butyl4-(tert-butoxycarbonylamino)-4-(1-(4-chlorophenyl)-4-hydroxybutylcarbamoyl)piperidine-1-carboxylate

O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (5.24 g, 13.77 mmol) was added in one portion to1-(tert-butoxycarbonyl)-4-(tert-butoxycarbonylamino)piperidine-4-carboxylicacid (Intermediate 70) (4.31 g, 12.5 mmol),4-amino-4-(4-chlorophenyl)butan-1-ol (Intermediate 16) (2.5 g, 12.5mmol) and DIPEA (6.56 mL, 37.6 mmol) in DMA (62.6 mL) under nitrogen.The resulting solution was stirred at 60° C. for 3 hours and evaporatedto dryness. The crude reaction mixture was dissolved in 100 mL EtOAc andthen basified with saturated NaHCO₃ (150 mL) and the organic fractiondried over MgSO₄, filtered and evaporated to dryness. The organicfraction was purified by flash silica chromatography (eluent 0-10% 7Nammonia in MeOH/DCM). Pure fractions were then evaporated to affordtert-butyl4-(tert-butoxycarbonylamino)-4-(1-(4-chlorophenyl)-4-hydroxybutylcarbamoyl)piperidine-1-carboxylate (5.06 g, 77%) as an off-white solid.

¹H NMR (400.13 MHz, DMSO) δ 1.39 (18H, s), 1.42-1.50 (2H, sex),1.67-1.73 (2H, q), 1.74-1.82 (2H, m), 1.87 (2H, broad), 3.07 (2H,broad), 3.34-3.41 (2H, m), 3.53-3.61 (2H, t), 4.71-4.76 (1H, q), 6.79(3×change, broad), 7.29-7.34 (4H, m), 7.80-7.82 (exchange, d).

MS m/e MH⁺=527; HPLC t_(R)=2.48 min.

Intermediate 72:4-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)piperidine-4-carboxamide

Tert-butyl4-(tert-butoxycarbonylamino)-4-(1-(4-chlorophenyl)-4-hydroxybutylcarbamoyl)piperidine-1-carboxylate(Intermediate 71) (1.98 g, 3.76 mmol) was stirred in THF (18.82 mL) and4M hydrogen chloride (14.11 mL, 56.46 mmol) in dioxane was added withstirring. The resulting solution was stirred at ambient temperatureovernight. The crude mixture was purified by SCX chromatography (eluent20% 7N ammonia/MeOH in DCM). Pure fractions were evaporated to drynessto afford4-amino-N-(1-(4-chlorophenyl)-4-hydroxybutyl)piperidine-4-carboxamide(1.04 g, 85%), as a colourless, transparent gum.

¹H NMR (400.13 MHz) δ 1.16-1.27 (2H, dd), 1.29-1.51 (2H, sepd),1.70-1.76 (2H, q), 1.75-1.89 (2H, m), 2.65-2.72 (2H, m), 2.74-2.83 (2H,dq), 3.37-3.39 (2H, m), 4.39 (exchange, broad) 4.70-4.76 (1H, q),7.32-7.35 (2H, d), 7.36-7.39 (2H, d), 8.25-8.27 (exchange, d)

MS m/e MH⁺=326; HPLC t_(R)=1.70 min.

Intermediate 73: tert-butyl 2-amino-1-(4-chlorophenyl)ethylcarbamate

A solution of 2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)ethylmethanesulfonate (Intermediate 23) (535 mg, 1.53 mmol) in DMF (8 mL) wastreated with sodium azide (199 mg, 3.06 mmol) and the mixture was heatedat 80° C. for 1 hour. The mixture was cooled and allowed to stir at roomtemperature overnight. The solution was partitioned between ethylacetate and water. The organic layer was washed twice with water thendried with magnesium sulfate, filtered and concentrated until the finalvolume was approximately 5 mL. Ethanol (20 mL) and 10% palladium oncarbon (75 mg, 0.07 mmol) were added. The resulting suspension wasstirred under an atmosphere of hydrogen at ambient pressure andtemperature for 1 hour. The mixture was filtered and the filtrate wasconcentrated under reduced pressure to give tert-butyl2-amino-1-(4-chlorophenyl)ethylcarbamate as a gum (410 mg, 99%).

MS m/e MH⁺=271

Intermediate 74: tert-butyl1-(4-chlorophenyl)-2-(methylsulfonamido)ethylcarbamate

A solution of tert-butyl 2-amino-1-(4-chlorophenyl)ethylcarbamate(Intermediate 73) (220 mg, 0.81 mmol) and N-ethyldiisopropylamine (0.281mL, 1.63 mmol) in THF (5 mL) was treated with methanesulfonyl chloride(0.075 mL, 0.98 mmol). The resulting solution was stirred at ambienttemperature for 2 hours. The mixture was partitioned between DCM andsodium bicarbonate solution. The organic layer was concentrated and theresidue was purified by flash column chromatography on silica usinggradient elution (10% ethyl acetate/DCM to 30% ethyl acetate/DCM). Thedesired product, tert-butyl1-(4-chlorophenyl)-2-(methylsulfonamido)ethylcarbamate, was thusisolated as a colourless solid (154 mg, 54.3%).

¹H NMR (399.9 MHz, CDCl₃) δ 1.43 (9H, s), 2.92 (3H, s), 3.38-3.52 (2H,m), 4.68-4.84 (2H, m), 5.20-5.28 (1H, m), 7.23 (2H, d), 7.35 (2H, d)

MS m/e (M−H)−=347

Intermediate 75: N-(2-Amino-2-(4-chlorophenyl)ethyl)methanesulfonamide

tert-butyl 1-(4-chlorophenyl)-2-(methylsulfonamido)ethylcarbamate(Intermediate 74) (151 mg, 0.43 mmol) was treated with trifluoroaceticacid (2 mL). The solution was stirred for 1 hour at room temperature.The mixture was concentrated under reduced pressure. The crude productwas purified by ion exchange chromatography, using an SCX column. Theresidue was loaded onto the column in methanol and washed with methanol.The desired product was eluted from the column using 2M ammonia inmethanol and pure fractions were evaporated to dryness to affordN-(2-amino-2-(4-chlorophenyl)ethyl)methanesulfonamide (93 mg, 86%) as acolourless crystalline solid.

¹H NMR (399.9 MHz, CDCl₃) δ 2.89 (3H, s), 3.17 (1H, dd), 3.33 (1H, dd),4.12 (1H, dd), 4.74 (1H, br, s), 7.29 (2H, d), 7.34 (2H, d)

MS m/e (M−H)−=247

Intermediate 76: tert-butyl4-(1-(4-chlorophenyl)-2-(methylsulfonamido)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (88 mg, 0.23 mmol) was added in one portion to astirred solution of4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (79 mg, 0.22 mmol) and N-ethyldiisopropylamine(0.046 mL, 0.26 mmol) in NMP (3 mL). The mixture was treated withN-(2-amino-2-(4-chlorophenyl)ethyl)methanesulfonamide (Intermediate 75)(60 mg, 0.24 mmol). The solution was stirred for 65 hours at roomtemperature. The mixture was partitioned between ethyl acetate andwater. The organic layer was washed twice with water. The organicsolution was concentrated under reduced pressure. The residue waspurified by flash column chromatography on silica using gradient elution(1% methanol/DCM to 15% methanol/DCM). Product containing fractions werecombined to give tert-butyl4-(1-(4-chlorophenyl)-2-(methylsulfonamido)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(103 mg, 79%) as colourless solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.41 (9H, s), 2.00 (4H, br, s), 2.81 (3H,s), 3.52-3.69 (2H, m), 4.18-4.27 (2H, m), 4.96 (1H, q), 6.60 (1H, s),6.97 (1H, br, s), 7.10-7.22 (2H, m), 7.37 (4H, s), 8.01 (1H, d), 8.14(1H, s), 11.65 (1H, s)

MS m/e MH⁺=592

Intermediate 77: 2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)ethylethanethioate

A solution of 2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)ethylmethanesulfonate (Intermediate 23) (600 mg, 1.72 mmol) in DMF (10 mL)was treated with potassium thioacetate (392 mg, 3.43 mmol) and themixture was heated at 50° C. for 1 hour. The mixture was cooled andpartitioned between ethyl acetate and water. The organic layer waswashed twice with water then dried with magnesium sulfate, filtered andconcentrated to dryness. The residue was purified by flash columnchromatography on silica using gradient elution (10% ethylacetate/isohexane to 20% ethyl acetate/isohexane). The desired product,2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)ethyl ethanethioate (509mg, 90%), was thus isolated as a cream crystalline solid.

¹H NMR (399.9 MHz, CDCl₃) δ 1.40 (9H, s), 2.35 (3H, s), 3.15-3.28 (2H,m), 4.78 (1H, br, s), 5.07 (1H, br, s), 7.24 (2H, d), 7.31 (2H, d)

MS m/e (M−H—CH₃CO)—=286

Intermediate 78: tert-butyl1-(4-chlorophenyl)-2-(chlorosulfonyl)ethylcarbamate

N-Chlorosuccinimide (819 mg, 6.14 mmol) was added to a solution of 2Mhydrochloric acid (0.8 mL) in acetonitrile (10 mL). The reaction flaskwas cooled with an ice bath to 10° C. and2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)ethyl ethanethioate(Intermediate 77) (506 mg, 1.53 mmol) was added portionwise. The mixturewarmed during the addition and was stirred for 10 minutes at roomtemperature. The mixture was partitioned between ethyl acetate andwater. The organic layer was washed with brine, dried with magnesiumsulfate, filtered and concentrated to dryness. tert-butyl1-(4-chlorophenyl)-2-(chlorosulfonyl)ethylcarbamate (602 mg, quant.) wasthus obtained as a colourless solid.

¹H NMR (399.9 MHz, CDCl₃) δ 1.44 (9H, s), 2.77 (1H, s), 4.06 (1H, dd),4.36 (1H, br, s), 5.15-5.23 (1H, m), 5.29-5.37 (1H, m), 7.29 (2H, d),7.38 (2H, d)

Intermediate 79: tert-butyl 1-(4-chlorophenyl)-2-sulfamoylethylcarbamate

Ammonia (1.5 mL, 31.50 mmol) was added to a suspension of tert-butyl1-(4-chlorophenyl)-2-(chlorosulfonyl)ethylcarbamate (Intermediate 78)(0.542 g, 1.53 mmol) in acetonitrile (10 mL). The mixture was stirredfor 16 hours at room temperature. The mixture was partitioned betweenethyl acetate and water and the organic layer was washed with brine. Theorganic solution was dried with magnesium sulfate, filtered, andconcentrated under reduced pressure. The residue was purified by flashcolumn chromatography on silica using gradient elution (10% ethylacetate/DCM to 30% ethyl acetate/DCM). The desired product, tert-butyl1-(4-chlorophenyl)-2-sulfamoylethylcarbamate (0.351 g, 68.5%), was thusisolated as a colourless solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.36 (9H, s), 3.21-3.28 (1H, m), 3.47-3.56(1H, m), 5.02 (1H, br, s), 6.88 (2H, s), 7.35 (2H, d), 7.41 (2H, d),7.49-7.60 (1H, m)

MS m/e (M−H—)−=333

Intermediate 80: 2-amino-2-(4-chlorophenyl)ethanesulfonamide

Tert-butyl 1-(4-chlorophenyl)-2-sulfamoylethylcarbamate (Intermediate79) (325 mg, 0.97 mmol) was treated with trifluoroacetic acid (8 mL).The resulting solution was stirred for 15 minutes at room temperature.The mixture was concentrated under reduced pressure and the residue waspurified by ion exchange chromatography, using an SCX column. The columnwas washed with methanol and the desired product was eluted usingammonia in methanol (2M) and pure fractions were evaporated to drynessto afford 2-amino-2-(4-chlorophenyl)ethanesulfonamide (221 mg, 97%) as anear colourless solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 3.13-3.25 (2H, m), 4.39 (1H, dd),7.35-7.48 (4H, m)

MS m/e (M−H—)−=233

Intermediate 81: Tert-butyl4-(1-(4-chlorophenyl)-2-sulfamoylethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (128 mg, 0.34 mmol) was added in one portion to astirred solution of4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (115 mg, 0.32 mmol) and N-ethyldiisopropylamine(0.066 mL, 0.38 mmol) in NMP (2.5 mL). The mixture was treated with2-amino-2-(4-chlorophenyl)ethanesulfonamide (Intermediate 80) (75 mg,0.32 mmol). The solution was stirred for 1 hour at room temperature. Themixture was treated with water (5 mL) and the resulting colourlessprecipitate was isolated by filtration, washed with water and thenacetonitrile to give tert-butyl4-(1-(4-chlorophenyl)-2-sulfamoylethylcarbamoyl)-1-(7H-(157 mg, 85%) ascolourless solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.40 (9H, s), 1.90-2.10 (4H, m), 3.35-3.39(1H, m), 3.51-3.59 (1H, m), 3.62-3.71 (2H, m), 4.12-4.26 (2H, m),5.30-5.39 (1H, m), 6.59 (1H, s), 6.82 (2H, s), 7.09-7.18 (2H, m),7.30-7.43 (4H, m), 8.13 (1H, s), 8.19 (1H, d), 11.65 (1H, s)

MS m/e MH⁺=578

Intermediate 82: tert-butyl 2-acetamido-1-(4-chlorophenyl)ethylcarbamate

A solution of tert-butyl 2-amino-1-(4-chlorophenyl)ethylcarbamate(Intermediate 73) (0.208 g, 0.77 mmol) and N-ethyldiisopropylamine(0.266 mL, 1.54 mmol) in THF (5 mL) was treated with acetic anhydride(0.102 mL, 1.08 mmol). The resulting solution was stirred at ambienttemperature for 2 hours. The mixture was partitioned between DCM andsodium bicarbonate solution. The organic layer was concentrated and theresidue was purified by flash column chromatography on silica usinggradient elution (10% ethyl acetate/DCM to 40% ethyl acetate/DCM). Thedesired product, tert-butyl 2-acetamido-1-(4-chlorophenyl)ethylcarbamate(0.151 g, 62.7%), was thus isolated as a colourless solid.

¹H NMR (399.9 MHz, CDCl₃) δ 1.41 (9H, s), 1.98 (3H, s), 3.46-3.67 (2H,m), 4.74 (1H, br, s), 4.97-5.56 (1H, m), 5.89 (1H, br, s), 7.22 (2H, d),7.32 (2H, d)

MS m/e MH⁺=313

Intermediate 83: N-(2-amino-2-(4-chlorophenyl)ethyl)acetamide

Tert-butyl 2-acetamido-1-(4-chlorophenyl)ethylcarbamate (Intermediate82) (148 mg, 0.47 mmol) was treated with trifluoroacetic acid (2 mL).The solution was stirred for 1 hours at room temperature. The mixturewas concentrated under reduced pressure. The crude product was purifiedby ion exchange chromatography, using an SCX column. The residue wasloaded onto the column in methanol and washed with methanol. The desiredproduct was eluted from the column using 2M ammonia in methanol and purefractions were evaporated to dryness to affordN-(2-amino-2-(4-chlorophenyl)ethyl)acetamide (98 mg, 97%) as a paleyellow crystalline solid.

¹H NMR (399.9 MHz, CDCl₃) δ 1.61 (2H, br, s), 1.97 (3H, s), 3.28-3.37(1H, m), 3.44-3.52 (1H, m), 4.05-4.11 (1H, m), 5.78 (1H, br, s),7.28-7.36 (4H, m)

MS m/e MH⁺=213

Intermediate 84: tert-butyl4-(2-acetamido-1-(4-chlorophenyl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (89 mg, 0.23 mmol) was added in one portion to astirred solution of4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (80 mg, 0.22 mmol) and N-ethyldiisopropylamine(0.046 mL, 0.27 mmol) in NMP (3 mL). The mixture was treated withN-(2-amino-2-(4-chlorophenyl)ethyl)acetamide (Intermediate 83) (52 mg,0.24 mmol). The solution was stirred for 65 hours at room temperature.The mixture was partitioned between ethyl acetate and water. The organiclayer was washed twice with water. The organic solution was concentratedunder reduced pressure. The residue was purified by flash columnchromatography on silica using gradient elution (1% methanol/DCM to 15%methanol/DCM). Product containing fractions were combined to givetert-butyl4-(2-acetamido-1-(4-chlorophenyl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(100 mg, 81%) as colourless solid.

¹H NMR (399.9 MHz, DMSO-d6) δ 1.42 (9H, s), 1.78 (3H, s), 1.98 (4H, br,s), 3.36-3.43 (1H, m), 3.50-3.68 (2H, m), 4.18-4.28 (2H, m), 4.85-4.93(1H, m), 6.58-6.61 (1H, m), 7.14-7.23 (2H, m), 7.30-7.37 (4H, m), 7.84(1H, br, s), 8.12-8.17 (2H, m), 11.65 (1H, br, s)

MS m/e MH⁺=556

Intermediate 85: 2-methyl-1-trityl-1H-imidazole

Triethylamine (11.54 mL, 82.82 mmol) was added dropwise to2-methyl-1H-imidazole (4.0 g, 48.72 mmol) and chlorotriphenylmethane(14.94 g, 53.59 mmol) in acetonitrile (120 mL) at room temperature overa period of 20 minutes under nitrogen. The resulting suspension wasstirred at room temperature for 18 hours. Water (120 mL) and i-hexane(12 mL) were added and the slurry stirred for 30 minutes beforefiltering. The filtered solid was washed with water (3×15 mL) and driedunder vacuum to afford 2-methyl-1-trityl-1H-imidazole (15.41 g, 97%) asa cream solid which was used without further purification.

1H NMR (399.902 MHz, CDCl₃) δ 1.65 (3H, s), 6.70 (1H, d), 6.90 (1H, d),7.11-7.16 (6H, m), 7.29-7.35 (9H, m);

m/z (ESI+) (M+H)+=325; HPLC tR=2.88 min.

Intermediate 86: 1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethanol

Butyllithium (1.6M in hexanes) (4.24 mL, 6.78 mmol) was added dropwiseto 2-methyl-1-trityl-1H-imidazole (Intermediate 85) (2.0 g, 6.16 mmol)in THF (30 mL) cooled to −78° C. over a period of 20 minutes undernitrogen. The resulting dark red solution was stirred at −78° C. for 30minutes then a solution of 4-Chlorobenzaldehyde (0.867 g, 6.16 mmol) inTHF (10 mL) added dropwise. The reaction was allowed to warm slowly to0° C. then quenched with saturated NH₄C₁ (50 mL) and extracted with TBME(2×). The combined extracts were washed with brine, dried over MgSO₄,concentrated by evaporation then purified by flash silicachromatography, elution gradient 20 to 30% EtOAc in isohexane. Purefractions were evaporated to dryness to afford1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethanol (1.810 g, 63.1%)as a colourless solid. 1H NMR (399.902 MHz, DMSO) δ 2.05 (2H, d),4.59-4.63 (1H, m), 5.99 (1H, d), 6.66 (1H, d), 6.96-7.02 (9H, m), 7.26(2H, d), 7.37-7.40 (9H, m);

m/z (ESI+) (M+H)+=465; HPLC tR=3.54 min.

Intermediate 87:2-(1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethyl)isoindoline-1,3-dione

Di-tert-butyl azodicarboxylate (0.808 g, 3.51 mmol) was addedportionwise to 1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethanol(Intermediate 86) (1.36 g, 2.92 mmol), phthalimide (0.473 g, 3.22 mmol)and triphenylphosphine (0.921 g, 3.51 mmol) in THF (25 mL) at roomtemperature over a period of 15 minutes. The resulting solution wasstirred at room temperature for 60 minutes then concentrated byevaporation and purified by flash silica chromatography, elutiongradient 20 to 40% EtOAc in isohexane. Pure fractions were evaporated todryness to afford2-(1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethyl)isoindoline-1,3-dione(1.78 g, 102%) as a colourless foam.

m/z (ESI+) (M+H)+=594; HPLC tR=2.80 min.

Intermediate 88:1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethanamine

2-(1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethyl)isoindoline-1,3-dione(Intermediate 87) (1.78 g, 3.00 mmol) and hydrazine hydrate (0.204 mL,4.19 mmol) in methanol (30 mL) were refluxed for 3 hours. The resultingclear solution was left to cool to room temperature overnight thenfiltered. The filtrate was applied to a 20 g SCX cartridge and elutedwith methanol followed by 2N NH₃(MeOH). Product-containing fractionswere combined and concentrated by evaporation then purified by flashsilica chromatography, elution gradient 2 to 5% MeOH in DCM. Purefractions were evaporated to dryness to afford1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethanamine (0.942 g,67.8%) as a colourless solid.

1H NMR (399.902 MHz, CDCl₃) δ 2.16 (2H, d), 4.01 (1H, t), 6.74 (1H, d),6.90 (2H, d), 6.99 (1H, d), 7.08-7.13 (7H, m), 7.29-7.32 (10H, m);

m/z (ESI+) (M+H)+=464; HPLC tR=3.13 min.

Intermediate 89: tert-butyl4-(1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (166 mg, 0.44 mmol) was added portionwise to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (150 mg, 0.42 mmol),1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethanamine (Intermediate88) (193 mg, 0.42 mmol) and N-ethyldiisopropylamine (0.087 mL, 0.50mmol) in DMF (2.0 mL) at room temperature. The resulting solution wasstirred at room temperature for 3 hours then quenched in water (10 mL)to give a pale yellow ppt. The precipitate was collected by filtration,washed with water and dried under vacuum to afford tert-butyl4-(1-(4-chlorophenyl)-2-(1-trityl-1H-imidazol-2-yl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(320 mg, 95%) as a cream solid, which was used without furtherpurification.

1H NMR (399.902 MHz, CDCl₃) δ 1.40 (9H, s), 2.05-2.29 (6H, m), 3.56-3.61(2H, m), 4.35-4.42 (2H, m), 5.33 (1H, s), 6.50 (1H, d), 6.78 (1H, m),6.93 (2H, d), 6.97-7.01 (7H, m), 7.13 (2H, d), 7.26-7.35 (10H, m), 8.27(1H, s), 9.25 (1H, br s), 10.22 (1H, br s).

m/z (ESI+) (M+H)+=807; HPLC tR=2.28 min.

Intermediate 90: 1-(4-Chlorophenyl)-2-(1H-pyrazol-1-yl)ethanone

A solution of pyrazole (3.40 g, 49.98 mmol) and2-bromo-4′-chloroacetophenone (11.44 g, 49 mmol) dissolved in DME (70mL) was stirred at room temperature under argon for 5 days. Diethylether was added and the precipitate was filtered, washed with diethylether and dried in vacuo overnight to give the crude product, 8.842 g ofpure white crystals. Concentrated aqueous ammonia (30%; 36 ml) was addedto a suspension of 8.76 g of the crude product in 7 ml of water. Thereaction was stirred for 40 min after which the yellow crystals werefiltered, washed with a little water and dried in vacuo to afford1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethanone (5.04 g, 22.84 mmol,46.6%) as a pale yellow solid. Reextraction of the washings gave furtherproduct. This was purified by flash chromatography on silica gel elutingwith 0 to 10% ethyl acetate in dichloromethane. The solvent wasevaporated to dryness to afford1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethanone (2.78 g, 12.60 mmol,25.7%) as a colorless crystalline solid. m/z (ESI+) (M+H)+=221.

1H NMR (500 MHz, DMSO-d6) δ 5.83 (2H, s), 6.61 (1H, dd), 7.48 (1H, d),7.66 (2H, d), 7.73 (1H, d), 8.04 (2H, d)

Intermediate 91: (Z) and(E)-1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethanone O-methyl oxime

Methoxylamine hydrochloride (0.668 g, 8.00 mmol), was added to a stirredsolution of 1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethanone (Intermediate90) (1.103 g, 5 mmol), at room temperature under argon. The resultingsolution was stirred at room temperature overnight. The pyridine wasevaporated in vacuo and the residual solid was triturated with asaturated aqueous solution of sodium hydrogencarbonate (50-100 ml). Thesolid was filtered, washed with water and dried to afford (Z) and(E)-1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethanone O-methyl oxime (1.080g, 87%) as a dark yellow solid. m/z (ESI+) (M+H)+=250.

1H NMR (500 MHz, DMSO-d6) δ 3.99 (3H, s), 5.44 (2H, s), 6.19 (1H, dd),7.36 (1H, d), 7.42 (2H, d), 7.65 (2H, d), 7.71 (1H, d)

Intermediate 92: 1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethanamine

Borane tetrahydrofuran complex (15.00 mL, 15.00 mmol) was added to astirred solution of (Z) and(E)-1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethanone O-methyl oxime(Intermediate 91) (0.749 g, 3 mmol) dissolved in THF (30 mL) at roomtemperature under argon. The resulting solution was stirred under refluxfor 3 hours. The mixture was cooled in an ice/water bath and water (25ml) was carefully added followed by 20% NaOH (25 ml). The resultingbiphasic mixture was refluxed overnight with vigorous magnetic stirringand allowed to cool. Diethyl ether was added and the layers wereseparated. The organic layer was further extracted with diethyl etherdried over MgSO₄, filtered and evaporated to give the crude product (741mg, 55%). This was used directly in the following reaction.

1H NMR (500 MHz, CDCl₃) δ 4.23 (1H, dd), 4.42 (1H, dd), 4.61 (1H, dd),6.54 (1H, dd), 7.14 (2H, d), 7.34 (2H, d), 7.60 (1H, d), 7.88 (1H, d)

Intermediate 93: Tert-butyl4-(1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

HATU (0.209 g, 0.55 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (0.181 g, 0.50 mmol),1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethanamine (Intermediate 92)(0.202 g, 0.5 mmol) and N,N-diisopropylethylamine (0.174 mL, 1.00 mmol)in N-methyl-2-pyrrolidinone (3.0 mL) at room temperature under argon.The resulting solution was stirred overnight. The reaction mixture waspurified by preparative HPLC using a Waters X-Bridge reverse-phasecolumn (C-18, 5 microns silica, 19 mm diameter, 100 mm length, flow rateof 40 ml/minute) and decreasingly polar mixtures of water (containing0.2% ammonium carbonate) and acetonitrile as eluent. The fractionscontaining the desired compound were evaporated to dryness to affordtert-butyl4-(1-(4-chlorophenyl)-2-(1H-pyrazol-1-yl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(0.216 g, 76%) as a white crystalline solid.

m/z (ESI+) (M+H)+=565.

1H NMR (500 MHz, DMSO-d6) δ 1.40 (9H, s), 1.72 (1H, m), 1.87 (2H, m),2.00 (1H, m), 3.53 (1H, m), 3.52 (1H, m), 3.94 (1H, m), 4.13 (1H, m),4.38 (1H, dd), 4.74 (1H, dd), 5.30 (1H, dt), 6.14 (1H, dd), 6.57 (1H,d), 7.16 (1H, d), 7.25 (1H, s), 7.29 (4H, s), 7.43 (1H, d), 7.53 (1H,s), 8.12 (1H, s), 8.35 (1H, d), 11.67 (1H, s)

Intermediate 94: (Z)-1-(4-Chlorophenyl)-2-(thiazol-2-yl)ethenamine

A 2.5M solution of N-butyllithium (8.00 mL, 20.00 mmol) in hexanes, wasadded dropwise to 2-methylthiazole (1.983 g, 20 mmol) dissolved in THF(35.00 mL) over a period of 5 minutes under argon at −70° C. Theresulting pale yellow slurry was stirred at −70° C. for 2.5 hour.4-chlorobenzonitrile (2.75 g, 20.00 mmol) in THF (35.00 mL) was addeddropwise to the suspension, which was stirred at −70° C. for 90 minutesafter the end of the addition. The reaction mixture was allowed to warmto room temperature under stirring and darkened. The crude wasconcentrated to dryness and diluted with ethyl acetate then washed withwater, dried over magnesium sulphate and concentrated. The crude productwas purified by flash chromatography on silica gel eluting with 10 to100% ethyl acetate in dichloromethane. The solvent was evaporated todryness to afford (Z)-1-(4-chlorophenyl)-2-(thiazol-2-yl)ethenamine(1.466 g, 31.0%) as a pale yellow solid.

m/z (ESI+) (M+H)+=237

1H NMR (500 MHz, CDCl₃) δ 5.72 (1H, s), 6.54 (2H, m), 6.99 (1H, d), 7.38(d, 2H), 7.55 (2H, d), 7.68 (1H, d)

Intermediate 95: 1-(4-Chlorophenyl)-2-(thiazol-2-yl)ethanamine

A 2N solution of hydrochloric acid in MeOH at room temperature, wasadded dropwise to (Z)-1-(4-chlorophenyl)-2-(thiazol-2-yl)ethenamine(Intermediate 94) (1.45 g, 6.13 mmol) and traces of bromocresol greendissolved in MeOH (45 mL) until the solution turned from dark blue toorange. Sodium cyanoborohydride (0.481 g, 7.66 mmol) was added in oneportion to the stirred solution and orange color was maintained byadding dropwise the 2N solution of hydrochloric acid in methanol at roomtemperature. After 1 h stirring, solvent was removed under vacuum. thecrude was taken up into water, pH was adjusted around 2 by adding a fewdrops of the 2N solution of hydrochloric acid in methanol. The aqueousphase was washed with diethyl ether twice then pH was adjusted to 14 byadding a 6N aqueous solution of sodium hydroxide and the basic aqueousphase was extracted with diethyl ether twice. The organic phases werewashed with brine dried over magnesium sulphate and concentrated toafford 1-(4-chlorophenyl)-2-(thiazol-2-yl)ethanamine (1.310 g, 90%) as acolorless oil.

1H NMR (500 MHz, DMSO-d6) δ 2.10 (2H, m), 3.22 (1H, dd), 3.26 (1H, dd),4.22 (1H, t), 7.32 (2H, d), 7.37 (2H, d), 7.51 (1H, d), 7.66 (1H, d)

Intermediate 96: Tert-butyl4-(1-(4-chlorophenyl)-2-(thiazol-2-yl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

HATU (0.418 g, 1.10 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (0.361 g, 1.0 mmol),1-(4-chlorophenyl)-2-(thiazol-2-yl)ethanamine (Intermediate 95) (0.318g, 1.00 mmol) and N,N-diisopropylethylamine (0.348 mL, 2.00 mmol) inN-methyl-2-pyrrolidinone (5.0 mL) at room temperature under argon. Theresulting solution was stirred overnight. Et₂O was added to precipitatethe rude product, which was filtered, washed plentifully with Et₂O anddried in vacuo to give tert-butyl4-(1-(4-chlorophenyl)-2-(thiazol-2-yl)ethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(0.550 g, 94%).

1H NMR (500 MHz, DMSO-d6) δ 1.23 (9H, s), 1.74 (1H, m), 1.89 (2H, m),2.00 (1H, m), 3.10 (2H, dd), 3.52 (1H, m), 3.58 (1H, m), 4.00 (1H, m),4.19 (1H, m), 5.29 (1H, dt), 6.57 (1H, d), 7.17 (1H, d), 7.30 (2H, d),7.37 (2H, d), 7.54 (1H), d), 7.69 (1H, d), 8.13 (1H, s), 8.33 (1H, d),8.47 (1H, d), 11.68 (1H, s)

m/z (ESI+) (M+H)+=582

Intermediate 97: lithium3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate

Lithium hydroxide (1.436 mL, 2.87 mmol) 2N in water was added to astirred suspension of methyl3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate(Intermediate 35) (400 mg, 0.72 mmol) in THF (40 mL) at roomtemperature. The resulting suspension quickly went into solution and wasthen stirred at RT overnight. The following morning a white precipitatehad formed. This was filtered and dried in vacuo to give lithium3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate(396 mg, 100%).

1H NMR (500 MHz, DMSO-d6) δ 1.41 (9H, s), 1.89 (2H, m), 2.06 (2H, m),2.27 (2H, m), 3.30 (2H, m partially hidden by H2O), 4.33 (2H, m), 4.83(1H, m), 6.57 (1H, s), 7.14 (1H, s), 7.22 (2H, d), 7.32 (2H, d), 7.58(1H, m), 8.10 (1H, s), 10.02 (1H, s);

m/z (ESI−) (M-Li)−=541

Intermediate 98: tert-butyl4-(1-(4-chlorophenyl)-3-(dimethylamino)-3-oxopropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

HATU (152 mg, 0.40 mmol) was added in one portion to lithium3-(4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamido)-3-(4-chlorophenyl)propanoate(Intermediate 97) (200 mg, 0.36 mmol), dimethylamine hydrochloride (48.0mg, 0.59 mmol) and N,N-diisopropylethylamine (0.190 mL, 1.09 mmol) inN-methyl-2-pyrrolidinone (3.0 mL) at room temperature under argon. Theresulting solution was stirred overnight. Diethyl ether added toprecipitate the crude product which formed as a gum. This was rinsedwith Et₂O and dried to give the crude gum (770 mg) which was used insubsequent reactions without further purification.

Intermediate 99: tert-butyl 1-(4-chlorophenyl)-3-methoxypropylcarbamate

Sodium hydride (35.0 mg, 0.87 mmol) was added to tert-butyl1-(4-chlorophenyl)-3-hydroxypropylcarbamate (Intermediate 30) (200 mg,0.70 mmol) in THF (10 mL) at 0° C. under nitrogen. The mixture wasstirred at 0° C. for 15 minutes. Methyl iodide (0.044 mL, 0.70 mmol) wasadded dropwise, and the resulting suspension was stirred at 22° C. for 4hours. The reaction was quenched with KHSO₄ solution (1M, 0.5 mL) andwater (15 mL). The mixture was extracted with diethyl ether (3×20 mL);the combined extracts were washed with saturated brine (20 mL), driedover MgSO₄, filtered and evaporated to give crude product. The crudeproduct was purified by flash silica chromatography, elution gradient 20to 60% EtOAc in isohexane. Pure fractions were evaporated to dryness toafford tert-butyl 1-(4-chlorophenyl)-3-methoxypropylcarbamate (80 mg,38.1%) as a white solid.

1H NMR (399.902 MHz, CDCl₃) δ 1.40 (9H, s), 1.91 (1H, s), 2.01 (1H, s),3.30 (3H, s), 3.32 (2H, m), 4.79 (1H, br.s), 5.45 (1H, br.s), 7.20 (2H,d), 7.29 (2H, d); m/z (ESI+) (M+H)+=300, 302 (M+H+), 244.3, 246.3(M+H+−C₄H8); HPLC tR=2.58 min.

Intermediate 100: 1-(4-chlorophenyl)-3-methoxypropan-1-amine

Hydrogen chloride (4M in 1,4-dioxane, 0.667 mL, 2.67 mmol) was added totert-butyl 1-(4-chlorophenyl)-3-methoxypropylcarbamate (Intermediate 99)(80 mg, 0.27 mmol) in a mixture of DCM (5 mL) and methanol (2 mL) at 22°C. The resulting solution was stirred at 22° C. for 5 hours. The mixturewas concentrated and the residue was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 2M NH₃/MeOH and pure fractions were evaporated todryness to afford 1-(4-chlorophenyl)-3-methoxypropan-1-amine (47.0 mg,88%) as a colourless oil.

1H NMR (399.902 MHz, CDCl₃) δ 1.80-1.96 (2H, m), 3.31 (3H, s), 3.32 (1H,m), 3.43 (1H, m), 4.09 (1H, t), 7.27-7.31 (4H, m);

m/z (ESI+) (M+H)+=200.2, 202.3 (M+H+); HPLC tR=1.70 min.

Intermediate 101: tert-butyl4-(1-(4-chlorophenyl)-3-methoxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (55.2 mg, 0.15 mmol) was added to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (50 mg, 0.14 mmol) and N-ethyldiisopropylamine(0.029 mL, 0.17 mmol) in NMP (5 mL). The resulting solution was stirredat 50° C. for 10 minutes then cooled to ambient temperature.1-(4-Chlorophenyl)-3-methoxypropan-1-amine (Intermediate 100) (27.6 mg,0.14 mmol) was added as a solution in NMP (2 mL). The resulting mixturewas stirred at 22° C. for 16 hours. The reaction mixture was dilutedwith EtOAc (75 mL) and washed sequentially with water (6×75 mL), andsaturated brine (50 mL). The organic layer was dried over MgSO₄,filtered and evaporated to afford crude product. The crude product waspurified by flash silica chromatography, elution gradient 1 to 4% (10:1MeOH/conc. NH_(3 (aq))) in DCM. Pure fractions were evaporated todryness to afford tert-butyl4-(1-(4-chlorophenyl)-3-methoxypropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(52.0 mg, 69.2%) as a white solid.

1H NMR (399.902 MHz, DMSO) δ 1.42 (9H, s), 1.90 (2H, t), 1.92-2.05 (4H,m), 3.22 (3H, s), 3.29 (2H, t), 3.55 (2H, m), 4.24 (2H, m), 4.89 (1H,dt), 6.60 (1H, dd), 7.05 (1H, br.), 7.16 (1H, dd), 7.33 (4H, m), 7.99(1H, d), 8.13 (1H, s), 11.65 (1H, s);

m/z (ESI+) (M+H)+=543.4, 545.3; HPLC tR=2.31 min.

Intermediate 102:S-3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propyl ethanethioate

Potassium thioacetate (43.9 mg, 0.38 mmol) was added to3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propyl methanesulfonate(Intermediate 31) (70 mg, 0.19 mmol) in DMF (5 mL) at 22° C. Theresulting solution was stirred at 50° C. for 2 hours. The reactionmixture was diluted with EtOAc (50 mL) and washed sequentially with 20%saturated brine (4×50 mL) and saturated brine (25 mL). The organic layerwas dried over MgSO₄, filtered and evaporated to afford crude product.The crude product was purified by flash silica chromatography, elutionsolvent 15% EtOAc in isohexane. Pure fractions were evaporated todryness to affordS-3-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propyl ethanethioate(61.0 mg, 92%) as a white solid.

1H NMR (399.902 MHz, CDCl₃) δ 1.41 (9H, s), 1.98 (2H, dt), 2.32 (3H, s),2.83 (2H, m), 4.67 (1H, br.s), 4.88 (1H, br.s), 7.20 (2H, d), 7.30 (2H,d);

m/z (ESI+) (M+H+—C₄H8)=288.3, 290.3; HPLC tR=2.80 min.

Intermediate 103: tert-butyl1-(4-chlorophenyl)-3-sulfamoylpropylcarbamate

N-Chlorosuccinimide (90 mg, 0.67 mmol) was added to a mixture ofacetonitrile (3 mL) and 2M hydrochloric acid (0.1 mL) cooled to 10° C.S-3-(tert-Butoxycarbonylamino)-3-(4-chlorophenyl)propyl ethanethioate(Intermediate 102) (58 mg, 0.17 mmol) was added dropwise as a solutionin acetonitrile (2 mL) and the resulting solution was stirred at 22° C.for 1 hour. Concentrated aqueous ammonia solution (3 mL, 63.00 mmol) wasadded dropwise and the mixture stirred for 3 days. The mixture wasevaporated and the residue was partitioned between DCM (30 mL) andwater. The aqueous layer was extracted with DCM (30 mL) and the extractscombined with the organic layer. The combined organics were filteredthrough a phase-separating paper and evaporated. The residue waspurified by flash silica chromatography, elution gradient 30 to 80%EtOAc in isohexane. Product-containing fractions were evaporated todryness to afford tert-butyl1-(4-chlorophenyl)-3-sulfamoylpropylcarbamate (40.0 mg, 68.0%) as awhite solid.

1H NMR (399.902 MHz, CDCl₃) δ 1.42 (9H, s), 2.75-2.91 (2H, m), 3.11-3.17(2H, m), 4.74 (1H, br.s), 4.93 (1H, t), 7.23 (2H, d), 7.34 (2H, d);

m/z (ESI+) (M+H+—C4H8)=293.2, 295.2; HPLC tR=2.11 min.

Intermediate 104: 3-amino-3-(4-chlorophenyl)propane-1-sulfonamide

Hydrogen chloride (4M in dioxan, 1.0 mL, 4.00 mmol) was added totert-butyl 1-(4-chlorophenyl)-3-sulfamoylpropylcarbamate (Intermediate103) (38 mg, 0.11 mmol) in a mixture of DCM (5 mL) and methanol (2 mL)at 22° C. The resulting solution was stirred at 22° C. for 24 hours. Themixture was evaporated to dryness and the residue was purified by ionexchange chromatography, using an SCX column. The desired product waseluted from the column using 30% (2M NH₃ in MeOH) in DCM and purefractions were evaporated to dryness to afford3-amino-3-(4-chlorophenyl)propane-1-sulfonamide (20.00 mg, 73.8%) as acolourless gum.

m/z (ESI+) (M+H)+=249.2, 251.2; HPLC tR=1.32 min.

Intermediate 105: tert-butyl4-(1-(4-chlorophenyl)-3-sulfamoylpropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (30.9 mg, 0.08 mmol) was added to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (28 mg, 0.08 mmol) and N,N-diisopropylethylamine(0.016 mL, 0.09 mmol) in NMP (5 mL) at 22° C. The resulting suspensionwas stirred at 50° C. for 10 minutes. The mixture was cooled to ambienttemperature and 3-amino-3-(4-chlorophenyl)propane-1-sulfonamide(Intermediate 104) (19.27 mg, 0.08 mmol) was added as a solution in NMP(2 mL). The mixture was stirred at 22° C. for 3 days. The mixture wasdiluted with methanol and loaded onto an SCX column. The crude productwas purified by ion exchange chromatography; the desired product waseluted from the column using 2M NH₃/MeOH and pure fractions wereevaporated to dryness. The crude product was purified by flash silicachromatography, elution gradient 2 to 6% (10:1 MeOH/conc. NH_(3 (aq)))in DCM. Pure fractions were evaporated to dryness to afford tert-butyl4-(1-(4-chlorophenyl)-3-sulfamoylpropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(16.00 mg, 34.9%) as a white solid.

m/z (ESI+) (M+H)+=592.4, 594.3; HPLC tR=1.99 min.

Intermediate 106: tert-butyl3-amino-1-(4-chlorophenyl)-3-oxopropylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluoro-phosphate (466 mg, 1.23 mmol) was added in one portion totert-butyl-3-amino-3-(4-chlorophenyl) propanoic acid (245 mg, 0.82mmol), ammonium chloride (131 mg, 2.45 mmol) andN,N-diisopropylethylamine (0.811 mL, 4.90 mmol) in DMF (4 mL) at 20° C.under nitrogen. The resulting suspension was stirred at 20° C. for 18hours. The reaction mixture was evaporated to dryness and redissolved inEtOAc (25 mL), and washed sequentially with water (25 mL) and saturatedbrine (25 mL). The organic layer was dried over MgSO₄, filtered andevaporated to afford tert-butyl3-amino-1-(4-chlorophenyl)-3-oxopropylcarbamate (200 mg, 82%) as a whitesolid. m/z (ESI−) (M−H)−=297; HPLC tR=1.91 min.

Intermediate 107: 3-amino-3-(4-chlorophenyl)propanamide

HCl (4M) in 1, 4 dioxane (1.674 mL, 6.69 mmol) was added to tert-butyl3-amino-1-(4-chlorophenyl)-3-oxopropylcarbamate (Intermediate 106) (200mg, 0.67 mmol) in DCM (20 mL) at 20° C. The resulting solution wasstirred at 20° C. for 4 hours. The reaction mixture was evaporated andwas purified by ion exchange chromatography, using an SCX column. Thedesired product was eluted from the column using 7M NH₃/MeOH and purefractions were evaporated to dryness to afford3-amino-3-(4-chlorophenyl)propanamide (46.0 mg, 34.6%) as a white solid.

1H NMR (399.9 MHz, DMSO-d6) δ 0.99 (2H, t), 2.33-2.37 (2H, m), 3.38 (2H,s), 4.24 (1H, t), 7.35-7.42 (4H, m),

m/z (ESI+) (M+H)+=199; HPLC tR=1.14 min.

Intermediate 108: tert-butyl4-(3-amino-1-(4-chlorophenyl)-3-oxopropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-n,n,n′,n′-tetramethyluroniumhexafluoro-phosphate (129 mg, 0.34 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (82 mg, 0.23 mmol) and N,N-diisopropylethylamine(0.112 mL, 0.68 mmol) in DMF (4 mL) at 20° C. under nitrogen. Theresulting solution was stirred at 20° C. for 5 minutes.3-amino-3-(4-chlorophenyl)propanamide (Intermediate 107) (45 mg, 0.23mmol) was then added to the reaction and stirred for 3 hours. Thereaction mixture was concentrated and diluted with EtOAc (20 mL) andwashed with water (20 mL). The organic layer was dried over MgSO₄,filtered and evaporated to afford tert-butyl4-(3-amino-1-(4-chlorophenyl)-3-oxopropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(99 mg, 100%).

m/z (ESI+) (M+H)+=542; HPLC tR=1.88 min.

Intermediate 109: tert-butylN-[1-(4-chlorophenyl)-3-(phenoxycarbonylamino)-propyl]carbamate

Phenyl chloroformate (0.132 mL, 1.05 mmol) was added dropwise totert-butyl 3-amino-1-(4-chlorophenyl)propylcarbamate (Intermediate 25)(300 mg, 1.05 mmol) and sodium bicarbonate (133 mg, 1.58 mmol) indioxane (10 mL) at 20° C. under nitrogen. The resulting solution wasstirred at 20° C. for 2 hours. The reaction mixture was concentrated anddiluted with EtOAc (20 mL) and washed with water (20 mL). The organiclayer was dried over MgSO₄, filtered and evaporated to afford tert-butylN-[1-(4-chlorophenyl)-3-(phenoxycarbonylamino)-propyl]carbamate (427 mg,100%) as a gum.

m/z (ESI+) (M+H)+=405; HPLC tR=2.85 min.

Intermediate 110: tert-butyl 1-(4-chlorophenyl)-3-ureidopropylcarbamate

Ammonium chloride (113 mg, 2.11 mmol) was added in one portion totert-butylN-[1-(4-chlorophenyl)-3-(phenoxycarbonylamino)propyl]carbamate(Intermediate 109) (427 mg, 1.05 mmol) and triethylamine (0.882 mL, 6.33mmol) in DMF (3 mL). The resulting solution was stirred at 0.352 molarfor 24 hours. The reaction mixture was concentrated and diluted withEtOAc (20 mL) and washed with water (20 mL). The organic layer was driedover MgSO₄, filtered and evaporated to afford tert-butyl1-(4-chlorophenyl)-3-ureidopropylcarbamate (346 mg, 100%),

m/z (ESI+) (M+H)+=328; HPLC tR=1.91 min.

Intermediate 111: 1-(3-amino-3-(4-chlorophenyl)propyl)urea

Tert-butyl 1-(4-chlorophenyl)-3-ureidopropylcarbamate (Intermediate 110)(346 mg, 1.06 mmol) was dissolved in TFA (3 mL) and stirred at 20° C.for 2 hours. The reaction was vacuumed to dryness and was purified byion exchange chromatography, using an SCX column. The desired productwas eluted from the column using 0.35M NH₃/MeOH and pure fractions wereevaporated to dryness to afford 1-(3-amino-3-(4-chlorophenyl)propyl)urea(151 mg, 62.8%) as a yellow gum.

1H NMR (399.9 MHz, DMSO-d6) δ 1.61 (2H, q), 2.98 (2H, m), 3.82 (1H, t),4.05 (2H, s), 5.38 (2H, s), 5.94 (1H, s), 7.37 (4H, m),

m/z (ESI+) (M+H)+=228; HPLC tR=1.25 min.

Intermediate 112: Tert-butyl4-(1-(4-chlorophenyl)-3-ureidopropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-n,n,n′,n′-tetramethyluroniumhexafluoro-phosphate (378 mg, 0.99 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (240 mg, 0.66 mmol) and N,N-diisopropylethylamine(0.329 mL, 1.99 mmol) in NMP (2 mL) at 20° C. under nitrogen. Theresulting solution was stirred at 20° C. for 5 minutes.1-(3-amino-3-(4-chlorophenyl)propyl)urea (Intermediate 111) (151 mg,0.66 mmol) in NMP (2 mL) was then added to the reaction and stirred for18 hours. The reaction mixture was diluted with EtOAc (20 mL) and washedwith water (20 mL). The organic layer was dried over MgSO₄, filtered andevaporated to afford tert-butyl4-(1-(4-chlorophenyl)-3-ureidopropylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate (379 mg, 100%) as a white solid. m/z (ESI+) (M+H)+=571; HPLCtR=1.88 min.

Intermediate 113: 3-amino-3-(4-chlorophenyl)propanenitrile

HCl (4M) in 1,4-dioxane (0.668 mL, 2.67 mmol) was added in one portionto tert-butyl 1-(4-chlorophenyl)-2-cyanoethylcarbamate (Intermediate 24)(150 mg, 0.53 mmol) in DCM (4 mL) at 20° C. The resulting solution wasstirred at 20° C. for 18 hours. The reaction mixture was evaporated toafford 3-amino-3-(4-chlorophenyl)propanenitrile (HCl salt) (115 mg, 99%)as a white solid.

Intermediate 114:(4-chlorophenyl)-2-cyanoethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluoro-phosphate (306 mg, 0.81 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (194 mg, 0.54 mmol) and N,N-diisopropylethylamine (0.533 mL, 3.22mmol) in NMP (3 mL) at 20° C. under nitrogen. The resulting solution wasstirred at 20° C. for 5 minutes.3-amino-3-(4-chlorophenyl)-propanenitrile (Intermediate 113) (97 mg,0.54 mmol) in NMP (3 mL) was then added to the reaction and stirred for1 hour. The reaction mixture was concentrated and diluted with EtOAc (50mL) and washed with water (50 mL). The organic layer was dried overMgSO₄, filtered and evaporated to afford(4-chlorophenyl)-2-cyanoethylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(0.281 g, 100%).

m/z (ESI+) (M+H)+=524; HPLC tR=2.19 min.

Intermediate 115: tert-butyl1-(4-chlorophenyl)-3-(methylsulfonamido)propylcarbamate

Methanesulfonyl chloride (0.082 mL, 1.05 mmol) was added dropwise totert-butyl 3-amino-1-(4-chlorophenyl)propylcarbamate (Intermediate 25)(300 mg, 1.05 mmol) and N,N-diisopropylethylamine (0.367 mL, 2.11 mmol)in DCM (4 mL) at 20° C. The resulting solution was stirred at 20° C. for18 hours. The reaction mixture was concentrated and diluted with Et₂O(25 mL) and washed with water (25 mL). The organic layer was dried overMgSO₄, filtered and evaporated to afford crude product. The crudeproduct was purified by flash silica chromatography, elution gradient 0to 20% EtOAc in DCM. Pure fractions were evaporated to dryness to affordtert-butyl 1-(4-chlorophenyl)-3-(methylsulfonamido)propylcarbamate (275mg, 71.9%) as a white solid.

1H NMR (399.9 MHz, DMSO-d6) δ 1.37 (9H, s), 1.76 (1H, m), 1.82-1.88 (1H,m), 2.87 (3H, s), 2.89-2.91 (2H, m), 4.58 (1H, d), 7.00 (1H, t), 7.32(2H, d), 7.39 (2H, d), 7.48 (1H, d),

m/z (ESI+) (M+H)+=361; HPLC tR=2.25 min.

Intermediate 116: N-(3-amino-3-(4-chlorophenyl)propyl)methanesulfonamide

TFA (4 mL) was added to tert-butyl1-(4-chlorophenyl)-3-(methylsulfonamido)propylcarbamate (Intermediate115) (275 mg, 0.76 mmol) and stirred at 20° C. for 2 hours. The reactionwas vacuumed to dryness. The crude product was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 7M NH₃/MeOH and pure fractions were evaporated todryness to afford N-(3-amino-3-(4-chlorophenyl)propyl)methanesulfonamide(113 mg, 56.7%) as a colourless gum.

1H NMR (399.9 MHz, DMSO-d6) δ 1.69-1.72 (2H, m), 2.87 (3H, s), 2.94-2.98(2H, m), 3.18-3.19 (1H, m), 3.87 (1H, t), 7.35-7.40 (4H, m),

m/z (ESI+) (M+H)+=262; HPLC tR=1.43 min.

Intermediate 117: tert-butyl4-(1-(4-chlorophenyl)-3-(methylsulfonamido)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate

O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluoro-phosphate (245 mg, 0.65 mmol) was added in one portion to4-(tert-butoxycarbonylamino)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxylicacid (Intermediate 1) (155 mg, 0.43 mmol) and N,N-diisopropylethylamine(0.213 mL, 1.29 mmol) in NMP (2 mL) at 20° C. under nitrogen. Theresulting solution was stirred at 20° C. for 5 minutes.N-(3-amino-3-(4-chlorophenyl)propyl)methanesulfonamide (Intermediate116) (113 mg, 0.43 mmol) in NMP (2 mL) was then added to the reactionand stirred for 18 hours. The reaction mixture was concentrated anddiluted with EtOAc (20 mL) and washed with water (20 mL). The organiclayer was dried over MgSO₄, filtered and evaporated to afford tert-butyl4-(1-(4-chlorophenyl)-3-(methylsulfonamido)propylcarbamoyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate(261 mg, 100%).

m/z (ESI+) (M+H)+=606; HPLC tR=2.09 min.

The biological effects of the compounds of Formula (I) may be tested asset out below.

-   -   In Vitro MDA-MB-468 Human Breast Adenocarcinoma GSK-3        Phosphorylation Assay

This assay determines the ability of test compounds to inhibitphosphorylation of Serine-9 residue in Glycogen Synthase Kinase-3beta(GSK-3(3) as a surrogate marker of cellular PKB (Akt) activity, asassessed using Acumen Explorer Fluorescent Plate-Reader technology. AMDA-MB-468 human breast adenocarcinoma cell line (LGC Promochem,Teddington, Middlesex, UK, Catalogue No. HTB-132) was routinelymaintained in Dulbecco's modified Eagle's growth medium (DMEM;Invitrogen Limited, Paisley, UK Catalogue No. 11966-025) containing 10%heat-inactivated foetal calf serum (FCS; Sigma, Poole, Dorset, UK,Catalogue No. F0392) and 1% L-glutamine (Gibco, Catalogue No. 25030-024)at 37° C. with 5% CO₂ up to a confluency of 70-90%.

For the phosphorylation assay, the cells were detached from the cultureflask using Trypsin-EDTA (Invitrogen Limited, Catalogue No. 25300-062)and seeded into the wells of a black transparent-bottom Corning CostarPolystyrene 96 well plate (Fisher Scientific UK, Loughborough,Leicestershire, UK; Catalogue No. 3904 and DPS-130-020K) at a density of5000 cells per well in 100 μl of complete growth media. The cells wereincubated overnight at 37° C. with 5% CO₂ to allow them to adhere.

On day 2, the cells were treated with test compounds and incubated for 2hours at 37° C. with 5% CO₂. Test compounds were prepared as 10 mM stocksolutions in DMSO and dosed directly to required concentration into testwells using non-contact (acoustic dispensing of multiple 2.5 nl dropletsdirectly into assay wells) ECHO dosing technology (Labcyte Inc.Sunnyvale, California, USA). Each plate contained control wells withouttest compound.

20 μl of fixing buffer (Phosphate Buffered Saline (PBS) containing 10%formaldehyde; Sigma; Catalogue No. F1635) was then added to each well togive a final well concentration 1.6%. Plates were then incubated for 30minutes at room temperature prior to the fixative being removed. Eachwell was washed once with 250 μl of PBS and then 50 μl PBS added to eachwell. PBS was then aspirated and cells permeabilised and blocked byincubating each well with 50 μl of permeabilisation/blocking buffer (PBScontaining 0.5% Tween 20 (Sigma; Catalogue No. P5927) and 5% Marvel MilkPowder (Andrews Pharmacy Ltd, Macclesfield, Cheshire, UK; Catalogue No.APC100199)) for 1 hour at room temperature prior to staining.

Following removal of Perm/Block buffer, 50 μl of primaryanti-phospho-GSK-3β antibody (Cell Signalling Technology (New EnglandBiolabs (Uk) Ltd.), Hitchin, Hertfordshire, UK; Catalogue No. 9336diluted 1:400 in Blocking buffer (PBS containing 5% Marvel and 0.05%Tween/Polysorbate 20) was added to each well and incubated overnight at4° C.

Each well was washed three times in 250 μl of wash buffer (PBScontaining 0.05% polysorbate 20), and cells incubated for 1 hour at roomtemperature with 50 μl of secondary fluorescently-labelled anti-rabbitAlexa Fluor 488 antibody (Molecular Probes, Invitrogen Limited,Catalogue No. A11008) diluted 1:750 in blocking buffer. Plates werewashed to three times in 250 μl of wash buffer and stored containing 50μl of PBS at 4° C. until required.

Plates were analysed using an Acumen Explorer Plate-reader to quantifylevel of fluorescent signal that represents quantity ofphosphorylated-GSK-3β. Active compounds caused a decrease inphospho-GSK-3β phosphorylation relative to the maximum (undosed) controlfor each assay, which is measured by the number of phosphorylatedobjects per well, and enabled potency of PKB (Akt) inhibitors to bedetermined.

IC₅₀ calculation—IC₅₀ is the concentration of compound required to give50% effect over the range of activity affected by the compound, betweenmaximum (no compound) and minimum (excess level of compound) responsecontrol data. IC₅₀ values were determined by fitting backgroundcorrected, dose response assay data to a 4 parameter logistic curve fitequation model with the minimum response set to zero. This was doneusing an in-house developed algorithm within the Origin graphingsoftware package (OriginLab Corporation, Northampton, MA, USA).

Examples of the invention were tested in the above assay and their meanIC₅₀ values calculated. These values are shown in Table G.

-   -   In Vitro AKT1 Kinase Assay

This assay detects inhibitors of AKT1 (PKBα) kinase activity usingCaliper LabChip LC3000. The Caliper off-chip incubation mobility shiftassay uses a microfluidic chip to measure the conversion of afluorescent labelled peptide to a phosphorylated product by a respectivekinase. The complete enzyme reaction is carried out in microtitre platesand then quenched. The resulting stopped solutions are serially ‘sipped’through a capillary onto the chip, where the peptide substrate andphosphorylated product are separated by electrophoresis. They are thendetected via laser-induced fluorescence. Substrate and product areseparated into two peaks by the application of a high electric field anddirectly detected using fluorescence. The signature of the fluorescentsignal reveals the extent of the reaction.

For Echo dosing the solvent was 100% DMSO. A master plate was preparedwith 40 ul of 10 mM stock from our Primary Liquid Store in quadrant 1 ofa Labcyte 384 well plate. A 1 in 100 dilution was made from quadrant 1into quadrant 2 by removing 0.4 ul and p) adding it to 39.6 ul of DMSO.Subsequent 1 in 100 dilutions were made into quadrant 3 from quadrant 2and quadrant 4 from quadrant 3.

Multiple 2.5 nl droplets were dispensed from each quadrant of the masterplate using ECHO dosing technology (Labcyte Inc. Sunnyvale, California,USA) to generate the dose range that was required in the test. The doserange most commonly used was as follows: 100 uM, 30 uM, 10 uM, 3 uM, 1uM, 0.3 uM, 0.1 uM, 0.03 uM, 0.01 uM, 0.003 uM, 0.001 uM, 0.0001 uM.Each well was backfilled with Dimethyl Sulphoxide (DMSO) to a totalvolume of 120 nl, such that when the enzyme and substrate mix was addedthe final DMSO concentration was 1%. DMSO was added to max control wellsas 120 nl, minimum control wells were treated with 120 nl of compound ata concentration that inhibited the enzyme activity 100%.

Following addition of compound or control to the assay plate, 6 μlpeptide mix containing 3 μM substrate (5-FAM-GRPRTSSFAEG-CONH2; CRB) and40 μM ATP in Kinase base buffer (100 mM Hepes pH 7.5, 0.015% Brij-35)and 6 μl enzyme mix containing 8 nM AKT1/PKBα active enzyme (UpstateBiotechnology, Cat No. 14-276), 8 mM DTT and 20 mM MgCl₂ in kinase basebuffer was added. All buffers were made up with 18MΩ water. The plateswere sealed and incubated at room temperature for 50 minutes. Thereaction was stopped by the addition of 10 μl stop buffer (100 mM HepespH 7.5, 0.015% Brij-35 solution, 0.1% coating reagent #3, 40 mM EDTA, 5%DMSO) to each well (N.B. plates can be frozen after stopping and readlater). The plates were then analysed using the Caliper LabChip LC3000Drug Discovery System (Caliper Life Sciences, 1 Wellfield, PrestonBrook, Runcorn, WA7 3AZ) using the following separation conditions; −1.8PSI, −500 upstream voltage, −1700 downstream voltage, sample sip time of0.2 sec, post sample sip time of 30 sec and a final delay of 120 sec.Integration of the substrate and product peaks was carried out usingCaliper LabChip software and IC₅₀ curves were calculated using Origin™software (OriginLab Corporation, Northampton, MA, USA). Examples of theinvention were tested in the in vitro AKT1 enzyme assay and the meanIC₅₀ values obtained are presented in Table G.

TABLE G Cellular In vitro Example PKB Mean PKBα Mean Number IC₅₀ (μM)IC₅₀ (μM)  1   0.2 0.0076  2   0.13 0.0038  3   0.14 0.0067  3A, 3B  0.11, >3.10 0.0026, 0.27  4   0.23 0.005  5   0.27 0.0097  6   0.020.0012  7   0.14 0.0061  7A, 7B   0.10, >3.10 0.0045, 0.24  8   0.30.0071  9   0.09 0.0032 10   0.25 0.0042 11   0.1 0.002 11A, 11B   0.11,0.48 0.0028, 0.021 12   0.1 0.0027 13   0.09 0.0042 14   0.94 0.00315 >3.10 0.093 16   4.6 0.28 17   0.24 0.012 18   1.4 0.016 19   0.260.011 20   0.98 0.02* 21 >3.10 0.059* 22   0.17 0.0075 23   0.08 0.001324   0.06 0.0022 25   0.13 0.0029 26   0.09 0.0056 27   0.37 0.016* 28  0.47 0.0045 29   1.6 0.041 30   1.1 0.03 31   1.1 0.024 32   0.420.012 33   0.24 0.014 34   0.35 0.0092 35   0.28 0.004 36   0.38 0.01437   0.55 0.019 38   0.36 0.0054 39   1.9 0.05 40   1.9 0.032 41   0.540.016 42   4.3 0.032 43   0.87 0.031 44   1.8 0.0089 45   0.77 0.23 46  6.7 0.15 47   0.21 0.0036 48   0.16 0.0056 49   0.081 0.005 49A, 49B  0.077, 1.2 0.0033, 0.048 50   0.19 0.0043 51   0.31 0.01 52   0.0780.0037 53 >2.4 0.0063 54   0.18 0.0034 55   0.043 0.0039 56   1 0.016 57  0.49 0.012 58   2.9 0.031 59   0.84 0.0063 60   0.47 0.023 61   0.850.023 62   0.56 0.028 63   0.68 0.042 64   0.3 0.0095 65   0.87 0.006466   0.99 0.0066* 67   0.89 0.0077* 68   0.34 0.012 69   0.41 0.0069 * =tested once only

-   -   hERG Analysis    -   Cell Culture

The hERG-expressing Chinese hamster ovary K1 (CHO) cells described by(Persson, Carlsson, Duker, & Jacobson, 2005) were grown tosemi-confluence at 37° C. in a humidified environment (5% CO₂) in F-12Ham medium containing L-glutamine, 10% foetal calf serum (FCS) and 0.6mg/ml hygromycin (all Sigma-Aldrich). Prior to use, the monolayer waswashed using a pre-warmed (37° C.) 3 ml aliquot of Versene 1:5,000(Invitrogen). After aspiration of this solution the flask was incubatedat 37° C. in an incubator with a further 2 ml of Versene 1:5,000 for aperiod of 6 minutes. Cells were then detached from the bottom of theflask by gentle tapping and 10 ml of Dulbecco's Phosphate-BufferedSaline containing calcium (0.9 mM) and magnesium (0.5 mM) (PBS;Invitrogen) was then added to the flask and aspirated into a 15 mlcentrifuge tube prior to centrifugation (50 g, for 4 mins). Theresulting supernatant was discarded and the pellet gently re-suspendedin 3 ml of PBS. A 0.5 ml aliquot of cell suspension was removed and thenumber of viable cells (based on trypan blue exclusion) was determinedin an automated reader (Cedex; Innovatis) so that the cell re-suspensionvolume could be adjusted with PBS to give the desired final cellconcentration. It is the cell concentration at this point in the assaythat is quoted when referring to this parameter. CHO-Kv1.5 cells, whichwere used to adjust the voltage offset on IonWorks™ HT, were maintainedand prepared for use in the same way.

-   -   Electrophysiology

The principles and operation of this device have been described by(Schroeder, Neagle, Trezise, & Worley, 2003). Briefly, the technology isbased on a 384-well plate (PatchPlate™) in which a recording isattempted in each well by using suction to position and hold a cell on asmall hole separating two isolated fluid chambers. Once sealing hastaken place, the solution on the underside of the PatchPlate™ is changedto one containing amphotericin B. This permeablises the patch of cellmembrane covering the hole in each well and, in effect, allows aperforated, whole-cell patch clamp recording to be made.

A β-test IonWorks™ HT from Essen Instrument was used. There is nocapability to warm solutions in this device hence it was operated atroom temperature (−21° C.), as follows. The reservoir in the “Buffer”position was loaded with 4 ml of PBS and that in the “Cells” positionwith the CHO-hERG cell suspension described above. A 96-well plate(V-bottom, Greiner Bio-one) containing the compounds to be tested (at3-fold above their final test concentration) was placed in the “Plate 1”position and a PatchPlate™ was clamped into the PatchPlate™ station.Each compound plate was laid-out in 12 columns to enable ten, 8-pointconcentration-effect curves to be constructed; the remaining two columnson the plate were taken up with vehicle (final concentration 0.33%DMSO), to define the assay baseline, and a supra-maximal blockingconcentration of cisapride (final concentration 10 μM) to define the100% inhibition level. The fluidics-head (F-Head) of IonWorks™ HT thenadded 3.5 μl of PBS to each well of the PatchPlate™ and its undersidewas perfused with “internal” solution that had the following composition(in mM): K-Gluconate 100, KCl 40, MgCl₂ 3.2, EGTA 3 and HEPES 5 (allSigma-Aldrich; pH 7.25-7.30 using 10 M KOH). After priming andde-bubbling, the electronics-head (E-head) then moved round thePatchPlate™ performing a hole test (i.e. applying a voltage pulse todetermine whether the hole in each well was open). The F-head thendispensed 3.5 μl of the cell suspension described above into each wellof the PatchPlate™ and the cells were given 200 seconds to reach andseal to the hole in each well. Following this, the E-head moved roundthe PatchPlate™ to determine the seal resistance obtained in each well.Next, the solution on the underside of the PatchPlate™ was changed to“access” solution that had the following composition (in mM): KCl 140,EGTA 1, MgCl₂ 1 and HEPES 20 (pH 7.25-7.30 using 10 M KOH) plus 100μg/ml of amphotericin B (Sigma-Aldrich). After allowing 9 minutes forpatch perforation to take place, the E-head moved round the PatchPlate™48 wells at a time to obtain pre-compound hERG current measurements. TheF-head then added 3.5 μl of solution from each well of the compoundplate to 4 wells on the PatchPlate™ (the final DMSO concentration was0.33% in every well). This was achieved by moving from the most diluteto the most concentrated well of the compound plate to minimise theimpact of any compound carry-over. After approximately 3.5 minsincubation, the E-head then moved around all 384-wells of thePatchPlate™ to obtain post-compound hERG current measurements. In thisway, non-cumulative concentration-effect curves could be produced where,providing the acceptance criteria were achieved in a sufficientpercentage of wells (see below), the effect of each concentration oftest compound was based on recording from between 1 and 4 cells.

The pre- and post-compound hERG current was evoked by a single voltagepulse consisting of a 20 s period holding at −70 mV, a 160 ms step to−60 mV (to obtain an estimate of leak), a 100 ms step back to −70 mV, a1 s step to +40 mV, a 2 s step to −30 mV and finally a 500 ms step to−70 mV. In between the pre- and post-compound voltage pulses there wasno clamping of the membrane potential. Currents were leak-subtractedbased on the estimate of current evoked during the +10 mV step at thestart of the voltage pulse protocol. Any voltage offsets in IonWorks™ HTwere adjusted in one of two ways. When determining compound potency, adepolarising voltage ramp was applied to CHO-Kv1.5 cells and the voltagenoted at which there was an inflection point in the current trace (i.e.the point at which channel activation was seen with a ramp protocol).The voltage at which this occurred had previously been determined usingthe same voltage command in conventional electrophysiology and found tobe −15 mV (data not shown); thus an offset potential could be enteredinto the IonWorks™ HT software using this value as a reference point.When determining the basic electrophysiological properties of hERG, anyoffset was adjusted by determining the hERG tail current reversalpotential in IonWorks™ HT, comparing it with that found in conventionalelectrophysiology (−82 mV) and then making the necessary offsetadjustment in the IonWorks™ HT software. The current signal was sampledat 2.5 kHz.

Pre- and post-scan hERG current magnitude was measured automaticallyfrom the leak subtracted traces by the IonWorks™ HT software by taking a40 ms average of the current during the initial holding period at −70 mV(baseline current) and subtracting this from the peak of the tailcurrent response. The acceptance criteria for the currents evoked ineach well were: pre-scan seal resistance >60 MΩ, pre-scan hERG tailcurrent amplitude >150 pA; post-scan seal resistance >60 MΩ. The degreeof inhibition of the hERG current was assessed by dividing the post-scanhERG current by the respective pre-scan hERG current for each well.

REFERENCES

-   Persson, F., Carlsson, L., Duker, G., & Jacobson, I. (2005).    Blocking characteristics of hERG, hNav1.5, and hKvLQT1/hminK after    administration of the novel anti-arrhythmic compound AZD7009. J    Cardiovasc. Electrophysiol., 16, 329-341.-   Schroeder, K., Neagle, B., Trezise, D. J., & Worley, J. (2003).    Ionworks HT: a new high-throughput electrophysiology measurement    platform. J Biomol Screen., 8, 50-64.    Results of hERG Analysis

Example 9,(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide,was tested up to 100 μM according to the procedure described above and amean hERG IC₅₀ value of 177 μM was derived by extrapolation of thecurve.

In Vivo Experiments

Pharmacodynamic Analysis of PKB Substrate Proteins GSK3β and PRAS40 inResponse to(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide

2.5×10⁶ U87-MG cells (ATCC number HTB-14™)+50% matrigel were injecteds.c. (subcutaneously) into the flank of nude mice. When tumours reacheda volume of about 0.5 cm³ an acute dose of 150 or 300 mg/kg of(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(E9) was given p.o. (oral gavage). Animals were euthanised at thespecified timepoint and tumours dissected out and snap frozen in liquidnitrogen.

Ex-vivo tumour lysates were prepared in 10% triton-X-100 Tris Lysisbuffer containing protease and phosphatase inhibitors using ‘FastPrep24’ methodology (MP Biomedicals matrix #6910-500). Proteinconcentrations were estimated against a BSA standard curve using PierceBCA kit (#23225). pGSK3β was measured by western blotting and pPRAS40 bysolid phase sandwich ELISA (Biosource KH00421).

For western blotting, equivalent amounts of protein were resolved by4-12% gradient Bis-Tris polyacrylamide pre-cast gels (InvitrogenNP0323), transferred to Hybond C Extra nitrocellulose membranes(Invitrogen LC2001) and incubated with primary antiserum (pGSK3β ser9,Cell Signaling Technology #9336; total GSK3(3, BD Transduction Labs#610202) and subsequently with either horseradish peroxidase conjugatedanti-rabbit IgG (Cell Signaling Technology #7074) or anti-mouse IgG(Cell Signalling Technology #7076). Immunoreactive proteins weredetected by enhanced chemiluminescence (#34076 Pierce Supersignal Dura)and bands quantified with a ChemiGenius II (Syngene). Plotted valuesshow the percentage inhibition of pGSK3β compared to vehicle controls,following normalisation for total pGSK3β.

For ELISA (Enzyme linked Immuno Sorbent Assay), equivalent amounts ofprotein were added to a 96 well plate pre-coated with a ‘capture’pPRAS40 specific monoclonal antibody. A ‘detection’ antibody specificfor pPRAS40 (Thr 246) is then added followed by an HRP labelledanti-rabbit IgG antibody. The assay is then quantified using stabilisedTMB (tetramethylbenzidine) at 450 nm on a plate reader. Colour intensityis proportional to the concentration of pPRAS40 (Thr 246). The resultsare shown in FIG. 1 .

MCF-7 Anti-Tumour Study Using(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(E9)

Severe Combined Immuno-Deficient (SCID) mice were obtained from CharlesRiver Laboratories. The mice were housed and maintained in specific,pathogen-free conditions. For in vivo implant, cells were harvested fromT225 tissue culture flasks by a 2- to 5-minute treatment with 3× trypsin(Invitrogen) in EDTA solution followed by suspension in basic medium andthree washes in phosphate buffered saline (Invitrogen). Only single-cellsuspensions of greater than 90% viability, as determined by trypan blueexclusion, were used for injection. MCF-7 breast tumor cells (ATCCnumber HTB-22™) (5×10⁶ cells+50% Matrigel™) were injected subcutaneouslyin the left flank of the animal in a volume of 0.1 mL. Prior toimplantation of the MCF-7 cells, SCID mice were anaesthetized andimplanted with a 0.5 mg/21 day duration estrogen pellet. When mean tumorsize reached ˜0.3 cm³, the mice were randomized into control andtreatment groups. The treatment group received 300 mg/kg E9 solubilizedin a vehicle consisting of 10% (v/v) DMSO, 25% (w/v) Kleptose™ in water,by oral gavage. The control group received the vehicle alone, once dailyby oral gavage. Tumor volumes (measured by caliper) were recorded atintervals for the duration of the study. Mice were sacrificed by CO₂euthanasia. The tumor volume was calculated (taking length to be thelongest diameter across the tumor and width to be the correspondingperpendicular diameter using the formula:(length×width)×√(length×width)×(π/6). Growth inhibition from the startof treatment was assessed by comparison of the differences in tumorvolume between control and treated groups. Because the variance in meantumor volume data increases proportionally with volume (and is thereforedisproportionate between groups), data were log-transformed to removeany size dependency before statistical evaluation. Statisticalsignificance was evaluated using a one-tailed, two-sample t test. Theresults are shown in FIG. 2 .

Human Dose Prediction

Human dose prediction for clinical studies requires an estimate of humanpharmacokinetic (PK) parameters important for defining the eliminationT_(1/2) and the shape and magnitude of the plasma concentration vs timeprofile at a particular dose. These estimated parameters includeClearance, Volume of Distribution at Steady State (Vss), absorption rateconstant (Ka), bioavailability (F) and dosing frequency. Human dosepredictions also require some pharmacological evidence as to howexposure relates to efficacy (McGinnity, Collington, Austin & Riley,Current Drug Metabolism 2007 8 463-479).

For(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(E9), human clearance was estimated from intrinsic clearance (Clint)data determined in human hepatocytes corrected to in vivo clearance byincorporation of the well-stirred model (Riley, McGinnity & Austin, DrugMetabolism & Disposition 2005 33 (9) 1304-1311). Minor differences wereobserved in the plasma protein binding of the compound across rat, dogand human; consequently, observed rat and dog Vss values were correctedby the factor Fu, human/Fu (rat or dog) (where Fu=fraction unbound inplasma). Human Vss was estimated to be 3.3 L/kg using this approach.Fraction absorbed (Fabs) in humans was taken as the average across ratand dog. This Fabs parameter was then adjusted to bioavailability (F) bycorrection for hepatic extraction. The absorption rate (Ka) was set toensure that Tmax was 1 h for E9, a relatively conservative value giventhat observed pre-clinically, and should equally result in aconservative Cmax value for use in calculation of margins with respectto safety studies. The clinical dosing frequency desired was assumed tobe twice daily (BID).

Having defined the shape and magnitude of the human plasma concentrationvs time curve, the final step was to adjust the dose so that plasmaconcentrations are attained that are likely to achieve efficacy. Inmouse PD studies, inhibition of pGSK (a surrogate end-point foranti-tumour activity) has been observed when free plasma concentrationsexceed 1-fold IC₅₀ for each compound as measured in the cell assay ofAKT inhibitory activity, corrected for binding due to Foetal Calf Serumpresent in the assay. Consequently, the human PK model assumed that freeexposure to E9 must exceed the free PKB IC₅₀ to ensure clinicalefficacy.

Putting all of this data together,(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(E9) is predicted to require a human dose of about 7 mg/kg BID toachieve efficacy, with a half-life estimated to be around 6 hours.

LIST OF FIGURES

FIG. 1 : GSK3β and PRAS40 phosphorylation levels in samples taken fromU87-MG tumours grown in nude mice following an acute dose of 150 or 300mg/kg(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(E9), n=5 per timepoint.

FIG. 2 : Results of MCF-7 antitumour study in SCID mice using a oncedaily (QD) dose of 300 mg/kg(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide(E9).

The invention claimed is:
 1. A method of treating breast cancer,comprising: administering to a person in need thereof a therapeuticallyeffective amount of fulvestrant, and a therapeutically effective amountof a compound of Formula (I), or a pharmaceutically acceptable saltthereof:

wherein: Y represents CH or N; Z¹—Z² represents C(R⁶)═CH; where R⁶represents hydrogen, fluoro, chloro, bromo, cyano, methyl, ethyl,difluoromethyl, trifluoromethyl or cyclopropyl; n is 0, 1 or 2; R¹represents C₁₋₄alkyl, C₂₋₄alkenyl, C₂₋₄alkynyl, C₁₋₄alkoxy,C₁₋₄alkoxyC₁₋₄alkyl, fluoroC₁₋₄alkyl, aminoC₁₋₄alkyl, hydroxyC₁₋₄alkyl,cyano, cyanoC₁₋₄alkyl, C₃₋₆cycloalkyl, —(CH₂)_(p)NHCOCH₃,—(CH₂)_(p)NHSO₂CH₃, —(CH₂)_(p)NHCONH₂, —(CH₂)_(p)NHCONR²R³,—(CH₂)_(p)NR²R³, —(CH₂)_(p)SO₂NH₂, —(CH₂)_(p)SO₂NR²R³, —(CH₂)_(p)CONH₂,—(CH₂)_(p)CONR²R³ or —(CH₂)_(p)—R⁷; where p is 0, 1, 2 or 3; R²represents hydrogen or C₁₋₃alkyl; R³ represents C₁₋₃alkyl; and R⁷represents phenyl; R⁷ represents a 5 or 6 membered monocyclic heteroarylring which comprises 1, 2 or 3 heteroatoms selected from O, N or S; orR⁷ represents a monocyclic 4, 5, or 6 membered heterocyclic ring whichcomprises 1, 2 or 3 heteroatoms selected from O, N or S; wherein R⁷ isoptionally substituted by 1 or 2 substituents selected from C₁₋₄alkyl,trifluoromethyl, C₁₋₄alkoxy, fluoro, chloro, bromo, and cyano; R⁴represents hydrogen, fluoro, chloro, bromo, cyano or trifluoromethyl;and R⁵ represents hydrogen, fluoro, chloro or bromo.
 2. The methodaccording to claim 1, wherein the compound of Formula (I) has theconfiguration shown in Formula (IA):


3. The method according to claim 1, wherein Z¹—Z² represents CH═CH. 4.The method according to claim 1, wherein R¹ represents methoxyC₁₋₄alkyl,fluoroC₁₋₄alkyl, aminoC₁₋₄alkyl, hydroxyC₁₋₄alkyl, cyanoC₁₋₄alkyl,C₃₋₆cycloalkyl, —(CH₂)_(p)NHCOCH₃, —(CH₂)_(p)NHSO₂CH₃,—(CH₂)_(p)NHCONH₂, —(CH₂)_(p)NHCONR²R³, —(CH₂)_(p)NR²R³,—(CH₂)_(p)SO₂NH₂, —(CH₂)_(p)CONH₂, —(CH₂)_(p)CONR₂R³ or —(CH₂)_(p)—R⁷.5. The method according to claim 4 wherein R¹ represents —(CH₂)_(p)—R⁷,p represents 1, 2 or 3, R⁷ is selected from phenyl, piperidinyl,piperazinyl, pyrrolidinyl, morpholinyl, imidazolyl, isoxazolyl,pyrazolyl and thiazolyl and R⁷ is optionally substituted by a singlemethyl group.
 6. The method according to claim 1, wherein R¹ representshydroxyethyl.
 7. The method according to claim 1, wherein R⁴ representschloro or bromo.
 8. The method according to claim 1, wherein R⁵represents hydrogen.
 9. The method according to claim 1, wherein n is 0or
 1. 10. A method of treating breast cancer, comprising: administeringto a person in need thereof a therapeutically effective amount offulvestrant, and a therapeutically effective amount of(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide:

or a pharmaceutically acceptable salt thereof.
 11. A method of treatingbreast cancer, comprising: administering to a person in need thereof atherapeutically effective amount of fulvestrant, and a therapeuticallyeffective amount of(S)-4-amino-N-(1-(4-chlorophenyl)-3-hydroxypropyl)-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamide: